CRAFT Symposium Program - SM

VIRTUAL
SYMPOSIUM
2021
PROGRAM
AUGUST
25 – 27
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2021
Winners of Microfluidics Art Competition
First place: Rick Lu, PhD Candidate, Institute of Biomedical Engineering,

University of Toronto
Image description: Confocal microscopy image of a vascularized tissue in

the Biowire II system. In this image, GFP-HUVEC (green) was stained with
VE-cadherin (yellow) and DAPI (blue).
Second place: Siwan Park, PhD Candidate, Institute of Biomedical

Engineering, University of Toronto
Image description: Pseudo-coloured SEM image of a carbon black

particle deposited on ciliated epithelial cells cultured in an organ-
on-a-chip system. In this image, the particle is coloured grey, cilia
are coloured orange, and epithelial cells are coloured green.
Third place: Zongjie Wang, PhD Candidate, Edward S. Rogers Sr.

Department of Electrical & Computer Engineering, University of
Toronto
Image description: SEM image revealing the 3D-printed

microstructure of a microfluidic system which has been applied to
early cancer diagnosis and cell enrichment for immunotherapy.
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Sponsors
Faculty of Applied Science & Engineering, University of Toronto
Faculty of Arts & Science, University of Toronto
Leslie Dan Faculty of Pharmacy, University of Toronto
Temerty Faculty of Medicine, University of Toronto
National Science and Engineering Research Council of Canada
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Welcome Message
Dear symposium attendees and distinguished guests,
We are very pleased to welcome you to the 2nd installment of the Virtual Research Symposium of the Centre for
Research and Applications in Fluidic Technologies (CRAFT). Thank you all for spending these precious last days
of August with us!
This year, we will again take advantage of our virtually connected world. We are fortunate to have Matthias and
Daniela update us on their exciting work from both sides of the Röschtigraben in Switzerland, Yoon from Ulsan
in the Republic of Korea, Jason from Philadelphia and Claudia from Toronto. In addition, the virtual format
makes it easier to connect busy industry and government folks with eager students and postdoctoral fellows
during the Trainee Networking Event.
Please take advantage of the many opportunities to network and make new contacts with the researchers,
clinicians and companies attending the symposium. If you are a graduate student or postdoctoral researcher,
inquire about the soon-to-be posted CRAFT Research Officer positions - we look forward to receiving your
application! Please drop in for our CRAFT session on August 25 at 1pm to learn more about the newly funded
CRAFT projects and our soon-to-be open CRAFT Device and Tissue Foundries. If you are an industry
representative, let us know the opportunities you see to contribute to CRAFT projects.
CRAFT is a team sport and a bottom-up initiative that has been in the making for the past 15 years. We would
not exist without the great CRAFT staff; the keen support from U of T’s Central Administration through the
Strategic Initiative Fund, the Innovations and Partnerships Office and the Faculties of Applied Science &
Engineering, Arts & Science, Medicine and Pharmacy at U of T; the significant commitment by the NRC; many
undergraduate and graduate students, postdoctoral fellows, faculty members, government scientists, clinicians,
representatives of technology companies and alumni; and passionate administrators from across U of T, its
affiliated teaching hospitals and NRC. And a special thank you to the group of graduate students and
postdoctoral fellows from U of T, Ryerson University and Sunnybrook Hospital who helped put this event
together!
We hope you enjoy the symposium. We look forward to hearing from you, seeing you regularly in the CRAFT
Device and Tissue Foundries, and meeting you at the 3rd CRAFT Research Symposium next year.
Axel Guenther and Teodor Veres
Co-Directors, CRAFT
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Agenda
August 25: Biofabrication
8:50-9:00 Welcome remarks
• Teodor Veres (National Research Council Canada [NRC])
• Roman Szumski (VP Life Sciences, NRC)
9:00-9:45 Biofabrication keynote (live)

• Matthias Lütolf (Swiss Federal Institute of Technology in Lausanne) -
Engineering Epithelial Organoids-on-a-chip
• Chair: Paul Santerre (U of T)
9:45-10:00 – MOVEU MOVEMENT BREAK –
10:00-10:45 Selected biofabrication presentations* (10-min recorded presentations followed by

live Q&A with presenter)
• 10:00 Albert Gevorkian (Eugenia Kumacheva/Edmond Young lab) - Actuation
of three-dimensional-printed nanocolloidal hydrogel with structural anisotropy
• 10:15 Katya D'Costa (Paul Santerre lab) - Effects of sodium ascorbate on the
deposition of extracellular matrix proteins: Towards engineering a vascular
graft
• 10:30 Wuyang Gao (Guenther lab) - One-step formation of protein-based
tubular structures for functional devices and tissues
• Chair: Axel Guenther (U of T)
11:00-12:00 Poster session 1

• Presenters of posters 1 – 18 will be online, at their posters and ready to
answer questions in real time using chat/video tools available on their poster
page.
12:00-1:00 – LUNCH BREAK –
1:00-2:30 CRAFT session: updates, new facilities and projects

• 1:00 Updates from CRAFT (Axel Guenther)
• 1:15 Introduction to CRAFT Device Foundry
• 1:20 Introduction to NRC Bio-devices Fabrication Facility in Boucherville,
Quebec
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• 1:30 Each CRAFT Project Award recipient will give a 5-min presentation
followed by a 3-min Q&A
• Chair: Axel Guenther
2:30-2:45 – MOVEU MOVEMENT BREAK –
2:45-3:30 Fireside chat with former grad student become company founder
• Daniela Marino (CEO and founder, Cutiss AG)
• Chair: Sushant Singh (Guenther Lab)
3:30-4:30 Industry exhibit hall

• Company representatives will be online, at their booths and ready to answer
questions in real time using chat/video tools available on their booth page.
4:30-4:45 Awards announcement for poster session 1 and microfluidics image competition
• Axel Guenther
August 26: TOeP and Organ-on-a-Chip

9:00-9:45 Organ-on-a-chip keynote (live)
• Jason Burdick (University of Pennsylvania) - Microfluidics-generated microgels
for the fabrication of biomedical granular hydrogels
• Chair: Milica Radisic (U of T)
9:45-11:15 Trainee networking event

• This event hosted on the gather.town platform will give students and
postdoctoral fellows the opportunity to network and socialize with the VIP
guests listed below and other trainees.
• RSVP here: https://forms.gle/M2V2bbjU3M8n8sHq6
VIP guests
• Jason Burdick PhD, Organ-on-Chip Keynote Speaker; Robert D. Bent
Professor, University of Pennsylvania, USA
• Yoon-Kyoung Cho PhD, Diagnostics Keynote Speaker; Full Professor,
Biomedical Engineering, Ulsan National Institute for Science and Technology
(UNIST)
• Matthias Lütolf PhD, Organ-on-a-Chip Keynote Speaker; Professor, Institute of
Bioengineering,École Polytechnique Fédérale de Lausanne (EFPL),
Switzerland
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• Ana Ramos PhD, R&D Scientist, BenchSci – Toronto
• Lorenzo Gutierrez PhD, Site Director, StarFish Medical – Toronto
• May Tsai PhD, Director of Regulatory Affairs, Baylis Medical – Mississauga
• Mehdi Shahini PhD, Senior Project Manager, Abbott – Ottawa
• Leanna Levine PhD, President & CEO, ALine Inc – Los Angeles
• Farnoud Kazemzadeh PhD, Co-Founder and VP Engineering, Vital Bio –
Waterloo, Canada
• Byeong-Ui (Ben) Moon PhD, Research Officer, NRC – Boucherville
• Jimin Guo PhD, Research Officer, NRC – Boucherville
• Caroline Miville-Godin MASc, Technical Officer, Microfabrication Specialist,
NRC – Boucherville
• Jason Davis MSc, IP Advisor, NRC
• Priyum Koonjul PhD, Client Relationship Leader, Medical Devices, NRC
11:15-11:30 - BREAK -
11:30-12:15 Selected TOeP trainee presentations (10-min recorded presentations followed by 5-

min live Q&A)
• 11:30 Karl Wagner (Milica Radisic lab) - Investigating the role and mechanisms
of extracellular vesicle signalling in human cardiac tissue-on-a-chip models
• 11:45 Kayla Soon (Sara Nunes lab) - Development and characterization of
vessel arteriovenous malformations (AVM)-on-a-chip
• 12:00 Zhengkun Chen (Eugenia Kumacheva/Edmond Young labs) -
Microfluidic arrays of dermal spheroids: a throughput screening platform for
skincare products
• Chair: Alison McGuigan (U of T)
12:15-1:15 - LUNCH BREAK -
1:15-2:15 Poster session 2

• Presenters of posters 19– 31 will be online, at their posters and ready to
answer questions in real time using chat/video tools available on their poster
page.
2:15-3:00 Selected organ-on-a-chip presentations* (10-min recorded presentations followed by

5-min live Q&A)
• 2:15 Dawn Lin (Boyang Zhang/Shinichiro Ogawa labs) - A 3D vascularized liver
model on the IFlowPlateTM
• 2:30 Elena Refet-Mollof (Thomas Gervais lab) - HOnAChip (hypoxia on a chip):
Insights into treatment efficacy
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• 2:45 Qinghua Wu (Radisic lab) - High-throughput heart-on-a-chip platform for
studies of SARS-CoV-2-induced myocardial injury
• Chair: Edmond Young (U of T)
3:00-3:15 - BREAK -
3:15-4:30 TOeP business pitch competition (7-min recorded presentation followed by 7-min live
Q&A with judges)
• 3:20 Kayla Soon/ Safwat Khan/ Omar Mourad - Vasculochip: for the new
generation of vascular medicine
• 3:35 Katrina Vizely - TBD
• 3:50 Sina Kheiri/ Zhengkun Chen - Artificial Intelligence Microfluidic Platform
for Anticancer Drug (AIMPAD)
• 4:05 Scott Campbell - POoCHS: Polymers for organ-on-chip Healthcare
Solutions by PolyVert
• Chair: Christopher Dixon
• Judges:
• Wendy Naimark, Chief Technology Officer, Ripple Therapeutics
• John Reid, Executive Vice President Business Development, Nanology
Labs Inc
• Jack Lee, Director of Technology Commercialization, OBIO
4:30-4:45 Closing remarks and awards announcement for poster session 2 and TOeP trainee
presentations
• Milica Radisic
August 27: Diagnostics

9:00-9:45 Diagnostics keynote (live)
• Yoon-Kyoung Cho (Ulsan National Institute of Science and Technology) - Lab-
on-a-disc for the detection of rare cells, extracellular biomarkers and beyond
• Chair: Aaron Wheeler (U of T)
9:45-10:45 Selected diagnostics presentations* (10-min recorded presentations followed by 5-

min live Q&A)
• 9:45 Jenise Chen (Shana Kelley lab) - Disease monitoring of protease activity
using a reagentless molecular pendulum
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• 10:00 Jimin Guo (NRC) - Automation of next generation sequencing
workflows on a centrifugal microfluidics platform
• 10:15 Intesar Zalloum (Scott Tsai lab) - Controlled shrinkage of
microfluidically-generated microbubbles by tuning lipid concentration
• 10:30 Lidija Malic (NRC) - Rapid and automated detection of SARS-CoV-2
using isothermal nucleic acid amplification
• Chair: Scott Tsai (Ryerson University)
10:45-11:00 - BREAK –
11:00-12:00 Industry exhibit hall

• Company representatives will be online, at their booths and ready to answer
questions in real time using chat/video tools available on their booth page.
12:00-1:00 - LUNCH BREAK –
1:00-2:45 NRC Pandemic Response Challenge – Selected presentations* and clinical translation
keynote
• 1:00 Introductory remarks from Jean-Francois Houle (NRC)
• 1:15 Warren Chan (U of T) - Assay for instrument-less SARS-CoV-2 rapid
diagnostic for saliva
• 1:30 Alexandros Sklavounos (Aaron Wheeler lab) - Latex agglutination tests
for rapid, instrument-free COVID-19 diagnostics in saliva
• 1:45 Evan Amalfitano (Keith Pardee lab) - An automated microfluidics glucose
meter interface for point-of-care gene circuit-based diagnostics
• 2:00 CLINICAL TRANSLATION KEYNOTE (LIVE): Claudia dos Santos (St.
Michael’s Hospital, U of T) – The Pandemic Preparedness Program: Analytical
validation and translation of COVID detection and patient stratification
technologies for Critical Care
• Chair: Jean-Francois Houle
2:45-3:00 Closing remarks and awards announcement for business pitch session and CRAFT
People’s Choice Award
• Teodor Veres
* CRAFT People’s Choice Award: All selected biofabrication, organ-chip, diagnostics and
NRC Pandemic Response Challenge Program presentations are part of the CRAFT People’s
Choice Award competition. Be sure to vote for your favourite presentation by emailing the
presenter’s name to carrie.keiski@utoronto.ca by 2:15pm on Friday, August 27.
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ABSTRACTS
2021
KEYNOTE
PRESENTATIONS 10
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Matthias Lütolf is a Swiss biomedical engineer and a Full Professor at the Institute of
Bioengineering and the Institute of Chemical Sciences and Engineering of École
Polytechnique Fédérale de Lausanne (EPFL) where he heads the Laboratory of Stem Cell
Bioengineering. His research program combines stem cell biology with engineering
principles and quantitative thinking. His transdisciplinary research involves engineers,
chemists, physicists, cell and developmental biologists, and aims at developing
technologies that have true biological and medicinal function, and applicability. Matthias
focuses on the bioengineering of miniature tissues (organoids) that are generated from self-
organizing stem cells. He is also interested to gain insight into how complex three-
dimensional microenvironments control the behavior of stem cells. Matthias graduated in
Material Engineering at ETH Zurich in 1999 where he also carried out his PhD studies under
the supervision of Jeffrey Hubbell until 2003. His PhD thesis was awarded an ETH medal in
2004. From 2005 to 2007 he continued his research training with Helen Blau as a
Postdoctoral Fellow in Stem Cell Biology at Stanford University. In 2007 he joined EPFL to
found his own research group. He served as the Director of the Institute of Bioengineering
at EPFL from 2014 to 2018, and was made a Full Professor in the School of Life Sciences at
EPFL in 2018. He is an elected a member of the European Molecular Biology Organization
(EMBO).
Engineering epithelial organoids-on-a-chip

Matthias P. Lütolf, PhD
Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, School of Life Sciences and School of Engineering,
École Polytechnique Fédérale de Lausanne (EPFL)
Organoids form through poorly understood morphogenetic processes in which initially homogeneous ensembles of stem
cells spontaneously self-organize in suspension or within permissive three-dimensional extracellular matrices. Yet, the
absence of virtually any predefined patterning influences such as morphogen gradients or mechanical cues results in
extensive heterogeneity. Moreover, the current mismatch in shape, size and lifespan between native organs and their in
vitro counterparts hinders their wider applicability. In this talk I will discuss some of our ongoing efforts in developing
organoids-on-a-chip that are assembled within hydrogel devices by guiding cell-intrinsic self-patterning through
engineered stem cell microenvironments.
Scheduled for: August 25, 2021 from 9:00-9:45
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Jason A. Burdick is the Robert D. Bent Professor of Bioengineering at the University of

Pennsylvania. Jason’s research involves the development of hydrogels through techniques
such as photocrosslinking and self-assembly and their processing using approaches such as
electrospinning and 3D printing. The applications of his research range from controlling
stem cell differentiation through material cues to fabricating scaffolding for regenerative
medicine and tissue repair. Jason currently has over 270 peer-reviewed publications, he is
on the editorial boards of Journal of Biomedical Materials Research A, Biofabrication,
Bioengineering, and Advanced Healthcare Materials, and he is an Associate Editor for ACS
Biomaterials Science & Engineering. He has been recognized through numerous awards
such as a Packard Fellowship in Science and Engineering, an American Heart Association
Established Investigator Award, the Clemson Award for Basic Science through the Society
for Biomaterials, and the Acta Biomaterialia Silver Medal Award. Jason has also been
elected as a Fellow of the American Institute for Medical and Biological Engineering, to the
International College of Fellows of Biomaterials Science and Engineering, and as a Fellow
of the National Academy of Inventors. Lastly, he has founded several companies to
translate technology developed in his laboratory towards clinical application.
Microfluidics-generated microgels for the fabrication of biomedical

granular hydrogels
Jason A. Burdick, PhD
Department of Bioengineering, University of Pennsylvania
Hydrogels represent a class of biomaterials that have great promise for the repair of tissues, particularly due to our
ability to engineer their biophysical and biochemical properties. Hydrogels can provide instructive signals through
material properties alone (e.g., mechanics, degradation, structure) or through the delivery of therapeutics that can
influence tissue morphogenesis and repair. In recent years, we have transitioned from traditional hydrogels to granular
hydrogels that are comprised of smaller hydrogel units (i.e., microgels). Microgels can be readily fabricated through
microfluidics, with variations in microgel size, shape, and throughput based on device design. Granular hydrogels are
formed through the packing of microgels and can be designed to be injectable through shear-thinning behavior,
heterogeneous through microgel mixing, and porous to support cell invasion.
Here, I will give examples of the design and use of granular hydrogels based on hyaluronic acid for use as injectable
therapeutics for endogenous tissue repair or in 3D printing to fabricate hydrogel constructs. For cardiac therapeutics, we
injected heterogeneous granular hydrogels into the myocardium and showed selective microgel degradation to release
factors and introduce porosity for cellular ingrowth. In 3D printing, we jammed microgels to form shear-thinning and
self-healing hydrogels that could be printed either onto surfaces or within other hydrogels. These could be cell-laden or
stabilized where necessary with secondary crosslinking. Additionally, we designed conductive granular hydrogels,
through an in situ metal reduction process of silver onto microgels that resulted in high conductivity due to increased
surface area when compared to traditional hydrogels. Most recently, we have developed anisotropic microgels and
showed increased cellular invasion into formed granular hydrogels based on pore structure. Overall, the design of
granular hydrogels opens up new opportunities in the design of functional hydrogels for biomedical applications.
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Yoon-Kyoung Cho is currently a full professor in Biomedical Engineering at UNIST and a

group leader in the Center for Soft and Living Matter at the Institute for Basic Science (IBS),
Republic of Korea. She received her PhD in Materials Science and Engineering from the
University of Illinois at Urbana-Champaign in 1999, having obtained her MS and BS in
Chemical Engineering from POSTECH in 1994 and 1992, respectively. She worked as a
senior researcher (1999–2008) at Samsung Advanced Institute of Technology (SAIT), where
she participated in the development of in vitro diagnostic devices for biomedical
applications. Since she joined UNIST in 2008, she served as the chair of the school of Nano-
Bioscience and Chemical Engineering (2008–2014) and the school of Life Sciences (2014–
2015) and the director of World Class University (2009–2013) and BK21 (2013–2015)
programs. Yoon is a member of the National Academy of Engineering of Korea (NAEK), an
associate editor of the journal Lab on a Chip, a fellow of the Royal Society of Chemistry,
and a board of director of the Chemical and Biological Microsystems Society (CBMS). She
serves as a scientific advisory board member of Clinomics and Labspinner, S. Korea. Her
research interests range from basic sciences to translational research in microfluidics and
nanomedicine. Current research topics include a lab-on-a-disc for the detection of rare cells
and extracellular biomarkers, quantitative analysis of single cells, and system analysis of
cellular communication.
Lab-on-a-disc for the detection of rare cells and extracellular

biomarkers and beyond.
Yoon-Kyoung Cho, PhD
Center for Soft and Living Matter, Institute for Basic Science (IBS)
Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST)
Liquid biopsy, a noninvasive alternative to tissue biopsy, is attracting tremendous interest among cancer clinics because it
provides specific information on circulating biomarkers and helps doctors design the right treatments for the right patient
at the right time. However, the cost, complexity, and large sample volume required for efficient isolation of circulating
biomarkers, as well as the low sensitivity, are still major limitations. Here, we introduce our on-going research on “lab-on-
a-disc”, which applies centrifugal force to pump biological fluid such as whole blood or urine to analyze cancer-related
biomarkers. The lab-on-a-disc systems to isolate and detect liquid biopsy markers such as circulating tumor cells (CTCs),
circulating tumor DNA (ctDNA), and extracellular vesicles (EVs) are developed and tested with clinical samples such as
whole blood or urine from cancer patients. First, we will introduce the fluid-assisted separation technology (FAST), which
enables ultrafast, uniform, clog-free, and highly efficient filtration. Next, we will discuss our recent studies on EVs-based
cancer diagnostics inspired by widespread recognition that EVs may be pivotal in intercellular communication. We
examine clinical samples by analyzing multiple kinds of proteins and RNA of EVs from cancer patient’s plasma or urine
samples and show that the EVs could be a potentially useful biomarker in cancer diagnostics. In addition, we will
introduce a hand-powered centrifugal bacterial isolation device to determine the bacterial load which is designed for use
in resource-limited settings for ruling out unnecessary antibiotic usage. We believe that the innovative point-of-care
microfluidic tools can accelerate the translation towards use in real clinical settings to directly impact patient care.
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Dr. Claudia dos Santos is an Associate Professor of Medicine at the University of Toronto,
and a Scientist at the Keenan Research Centre for Biomedical Science of St. Michael’s
Hospital. She holds the Robert and Dorothy Pitts Chair in Acute Care and Emergency
Medicine. Her Clinical appointment is that of Staff Intensivist in the Medical Surgical ICU of
St Michael’s. She is also a Scientist of the Institute of Medical Sciences, cross appointed to
Laboratory Medicine and Pathobiology and a member of the Collaborative Program in
Genome Biology and Bioinformatics. She is an active member of the Canadian Critical Care
Translational Biology and Critical Care Trials Group. She is a proud recipient of the Parker
B. Francis Fellowship for Respiratory Research, Heart and Stroke Foundation and Canadian
Institutes of Health Research (CIHR) Fellow. Her major research interest is in acute lung
injury, which can be caused by either biomolecular or biophysical insults to the lungs, such
as an infection or the mechanical injury resulting from mechanical ventilation itself. Her lab
is dedicated to understanding the interaction between patients and the breathing machine
and finding new ways to identify individuals who are at higher risk for developing lung
injury, in addition to diagnosing, treating and monitoring improvement from injury.
The Pandemic Preparedness Program: Analytical validation and

translation of COVID detection and patient stratification technologies
for Critical Care
Claudia C. dos Santos, MSc, MD, FRCPC
Robert and Dorothy Pitts Chair in Acute Care and Emergency Medicine
Associate Professor of Medicine, University of Toronto
Clinician-Scientist, Scientist Keenan Center for Biomedical Research, St-Michael’s Hospital
Institute for Biomedical Engineering, Science & Technology (iBEST)
Background: Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-
CoV2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in
patient samples, but RNA extraction and conventional PCR constitutes a major bottleneck in current testing requiring
both expert personnel and time – causing important delays in care. Moreover, while pathogen diagnosis is critical,
patient outcomes vary depending upon the host’s immune response to the virus. Currently, there are no tests that
inform - at the point-of-need - how to risk stratify patients once they test positive for SARS-CoV2. Methodological
simplification and advancement in patient stratification could increase diagnostic availability and improve triage
efficiency, benefitting patient care and infection control. Advancement in precision-care will be especially important if
vaccines turn out not to be as efficacious as we hope, if new variants have different resistance and/or response patterns
and/or if resistance to existing therapeutics emerges.
We propose to innovate the care of the COVID19 patient, and other emerging future pathogens, by adapting the
existing NRC-Power-Blade digital droplet PCR (ddPCR) RNA detection technology to perform detection of host immune
response RNA-based markers in very small amounts of human biosamples. This proposal aims to combine the clinical
translational capacity of our team at St. Michael’s Hospital (University of Toronto), the bioinformatics expertise of our co-
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investigator Dr. Robert E. W. Hancock at Sepset Biosciences Inc. with the University of British Columbia, and the unique
microfluidic technology developed by the National Research Council (NRC) of Canada team led by Dr. T. Veres.
Aim 1: Adapt, develop and qualify the NRC PowerBlade to perform fast and high-quality RNA extraction from whole
blood and ddPCR-based multiplex assay to detect the human 8 gene signature able to risk stratify patients at the point-
of-need.
Aim 2: Integrate the workflow for the ddPCR-based CR-detection multiplex assay into the PowerBlade and build the
prototype POWER-PREDICT.
Aim 3: Perform analytical demonstration on the POWER-PREDICT prototype using 'real-world' patient samples with
known COVID status and benchmark sensitivity and specificity of human probe detection in patients with suspected
COVID prospectively.
Impact: Our proposal touches on all aspects of COVID19 management, from early diagnosis, prognosis, selection of
treatment, monitoring of therapeutic efficacy, and outcomes. The added-value of our project is in the enablement of the
research infrastructure and knowledge that will support host-stratification biomarkers development in a unique
translational environment for this as well as future pandemics impacting the life of patients in Canadians and all over the
world.
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MAIN PRESENTATIONS
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Actuation of three-dimensional-printed nanocolloidal hydrogel

with structural anisotropy*
Albert Gevorkian, Sofia M. Morozova, Sina Kheiri, Nancy Khuu, Heyu Chen, Edmond Young, Ning Yan, Eugenia
Kumacheva
Department of Chemistry, University of Toronto
Rationale and Objectives: Actuation occurs due to differential hydrogel swelling and is generally achieved by
modulating hydrogel composition. Hydrogels with a variation in composition across the sheet, in addition to
the top and bottom portions of the sheet can exhibit a different degree of stimulus-induced swelling or
contraction. Here, we report a different approach to hydrogel actuation using a 3D-printed single-layer
nanocolloidal hydrogel actuator that undergoes stimulus-triggered shape transformation solely due to structural
anisotropy.
Methods: Hydrogel sheets were composed of a suspension of cellulose nanocrystals(CNC),

methacryloyl(GelMA), and a photoinitiator. The ink was supplied to a custom-built microfluidic extruder and
printed on a cold stage and crosslinking, during which CNCs are aligned in the direction of extrusion. To induce
actuation, hydrogel sheets were submerged in water. To investigate the relation between the CNC alignment
direction and hydrogel sheet shape, hydrogel sheets were cut into different shapes and then submerged in
water. Lastly, to demonstrate programmable hydrogel morphing, selective crosslinking via photomasks and
GelMAs sol-gel transition were used to control the regions of CNC alignment in the hydrogel sheet.
Results: Shear induced CNC alignment can be preserved through a combination of cooling the ink below its
sol-gel transition and photo cross-linking. Aligned hydrogel sheets actuated via swelling induced formation of
rolls when placed in water. The extent of curvature was dependent on the concentration of cellulose
nanocrystals. We showed the relation between hydrogel sheet shape and the axis of CNC orientation. Sheets
whose long axis were aligned with the direction of CNC alignment, perpendicular, and diagnol to the direction
of CNC alignment all formed different shapes. Lastly, swelling induced hydrogel morphing was achieved by
selectively controlling the regions of local alignment, generating a veriety of shapes.
Conclusion and Significance: We show a new biocompatible, shear-thinning and self-healing nanocolloidal ink,
for the programmable actuation of single-layer hydrogels, solely from structural anisotropy. Multiple shapes
were generated by the same hydrogel actuator. Future work will focus on using this methodology to generate
materials for modeling structurally anisotropic biological tissues.
*Competing in CRAFT People’s Choice Award
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Effects of sodium ascorbate on the deposition of extracellular

matrix proteins - towards engineering a vascular graft*
Katya D'Costa, Nicole Machado, Craig A. Simmons, J. Paul Santerre
Institute of Biomedical Engineering, University of Toronto; Department of Pharmacology and Toxicology,

University of Toronto; Translational Biology and Engineering Program, Ted Rogers Centre for Heart Research;
Department of Mechanical and Industrial Engineering, University of Toronto; Faculty of Dentistry, University of
Toronto
Rationale and Objectives: Pulmonary hypertension is defined by stiffening of arteries in the lungs, ultimately
resulting in heart failure and death. Shunting oxygenated blood with vascular grafts could mitigate the need for
cardiopulmonary bypass. However, commercial grafts with small diameters suffer from low long-term patency
rates and poor mechanical compliance. The objective of this study was to generate a cell-seeded scaffold
system with a confluent sheet of vascular smooth muscle cells (VSMCs) that could recapitulate the architecture
of native arteries.
Methods: Adipose stromal cells (ASCs) were isolated from human abdominal adipose tissue and differentiated
into VSMC-like cells (ASC-VSMCs). Human monocytes were isolated from whole blood samples and co-seeded
through a two-stage process at a ratio of 4:1 (ASC-VSMC: monocyte) on nanofibrous (481±143 nm)
polyurethane scaffolds at a density of 70,000 cells/cm2. Cultures were supplemented with 100 µM of sodium
ascorbate which was previously observed to enhance elastin and glycosaminoglycan (GAG) production.
Immunostaining was carried out with established early, mid- and late-stage markers for VSMC maturity and
elastin. Biochemical assays were used to quantify ECM protein production in cultures over the duration of one
month.
Results: One week post culture, ASC-VSMCs were observed to form a confluent and aligned layer across the
nanofibrous sheet in the sodium ascorbate supplemented (SA+) culture; however, the cells lacking
supplementation (SA-) failed to achieve confluency. In contrast, staining carried out on the samples at two
weeks indicated both conditions had achieved confluence. The production of major ECM constituents including
collagen, elastin and GAGs were measurably enhanced at the four-week timepoint, with a proposed
phenotypic change occurring at week two. Additionally, cultures from the second week appeared to have more
elastin than the first week for both groups.
Conclusion and Significance: As demonstrated, electrospun scaffolds could be seeded in static culture with
primary monocytes and vascular smooth muscle cells derived from a human host. Given the favorable results, it
is anticipated that the biochemical factors from monocytes and applied mechanical stimuli will enable in vitro
vascular tissue engineering with a functional ECM.
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2021
One-step formation of protein-based tubular structures for

functional devices and tissues*
Wuyang Gao, Nima Vaezzadeh, Kelvin Chow, Haotian Chen, Patricia Lavender, Mark D. Jeronimo, Arianna
McAllister, Onofrio Laselva, Jia-Xin Jiang, Blair K. Gage, Shinichiro Ogawa, Arun Ramchandran, Christine E.
Bear, Gordon M. Keller, and Axel Günther
Dept. of Mechanical and Industrial Engineering; Inst. of Biomedical Engineering; The Hospital for Sick Children,
McEwen Stem Cell Institute, University Health Network, Toronto, Canada; Dept. of Chemical Engineering and
Applied Chemistry, University of Toronto
Rationale and Objectives: Tubular structures consisting of extracellular matrix (ECM) proteins and cells are basic
functional organ units in animals and humans. ECM protein solutions at low concentration are abundantly used
in 3D cell culture. However, their poor “printability” and slow gelation kinetics have made the direct extrusion
challenging. Here, we overcome this limitation and demonstrate the continuous, template-free conversion of
low-concentration collagen, elastin, and fibrinogen solutions into tubular structures of tailored properties and
composition.
Methods: The extrusion-based bioprinting approach is enabled by a microfabricated multi-layer PDMS

printhead where ECM protein solution is hydrodynamically focused to occupy an annulus between two buffer
solutions (PEG). Subsequently, solutions enter a downstream confinement (25 cm) accommodating the minute-
long residence times required for ECM protein solutions to gel. Custom designed tube hosting device with
individual perfusion, superfusion, and transmural pressure was built to characterize mechanical properties (e.g.,
Young's modulus, permeability, elastocapillary effect, etc.) of obtained tubular structures and demonstrate their
tissue-level function characteristics with residence cells.
Results: We have developed co-axial extrusion bioprinting approach for tubular structure from a variety of
natural ECM protein materials. Importantly, we demonstrated the formation of cell-laden collagen tubes using a
biocompatible, neutral pH collagen gelation approach, which has thus far remained a critical limitation of ECM
bioprinting. Hosted on a custom device, we demonstrated the obtained ECM tubular structures to assume
device level functional characteristics, collapse and re-opening of tubular structures, including building all-ECM
gate valves and their in-series configuration as peristaltic pumps. In addition, we have demonstrated functional
endothelial and epithelial barrier layers.
Conclusion and Significance: We have developed an extrusion bioprinting approach for tubular structure from a
variety of slow-gelling natural ECM protein materials. Obtained ECM tubular structures enable studies of
physiologically relevant elastocapillary effects and promises an in vitro avenue for systematically recapitulating a
variety of intact ductular tissues.
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SYMPOSIUM
2021
Investigating the role and mechanisms of extracellular vesicle

signalling in human cardiac tissue-on-a-chip models
Karl T. Wagner, Milica Radisic
Department of Chemical Engineering and Applied Chemistry, University of Toronto; Institute of Biomedical
Engineering, University of Toronto; Toronto General Research Institute, University Health Network; The Heart
and Stroke/Richard Lewar Centre of Excellence, Toronto
Rationale and Objectives: Heart diseases are the leading cause of death worldwide. To discover new
treatments, it is important to understand cardiac physiology at a cellular level. Studies indicate that cell-
signalling via release and uptake of extracellular vesicles (EVs) plays an integral role in both healthy and
diseased hearts. The “organ-on-a-chip” industry has pioneered fabrication of human tissue models, combining
cells and biomaterials to mimic organ function. These models represent a relevant, accessible approach for
investigating the role of EVs in the heart.
Methods: The “Biowire” model of cardiac tissue-on-a-chip will be used to investigate the role of EVs in cardiac
physiology and ischemic injury. Biowires fabricated from differentiated induced pluripotent stem cells (iPSC) will
be subjected to baseline functional analyses alongside compositional profiling of tissue-secreted EVs. A model
of ischemic injury will be established in Biowires, followed by EV profile comparison to healthy tissues. EVs
isolated from 2D cultures of iPSC, cardiomyocytes, cardiac fibroblasts, endothelial cells, and healthy Biowires
will be added separately to injured tissues, assessing functional response to different sources of EVs and
predicting their regenerative potential.
Results: EVs secreted by Biowire tissues were successfully isolated and characterized. Nanoparticle tracking
established the size distribution and concentration of secreted particles, while western blotting and
transmission electron microscopy confirmed the presence of EVs in isolates. Biowire tissues cultured in hypoxia
and ischemia-reperfusion simulating culture media displayed a measurable decrease in viability. Sequencing
and differential expression analysis of microRNA within EVs isolated from 2D cardiac cell cultures were used to
predict the potential utility and functional effects of applying different EV populations to the injured heart as a
regenerative therapeutic.
Conclusion and Significance: Adapting the physiologically relevant Biowire model will facilitate mechanistic
study of cardiac EVs at a level not previously possible. This knowledge is a step towards developing EV-based
therapies to regenerate damaged hearts that have the potential to revolutionize patient quality of life and
reduce the societal burden of heart disease.
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2021
Development and characterization of vessel arteriovenous

malformations (AVM)-on-a-chip
Kayla Soon, Ruilin Wu, Jason Fish and Sara S. Nunes
The Heart and Stroke/Richard Lewar Centre of Excellence, Toronto General Hospital Research Institute,
University Health Network, Institute of Biomedical Engineering, and Laboratory Medicine and Pathobiology,
Rationale and Objectives: Arteriovenous malformation is the abnormal formation of blood vessels characterized
by the direct connection between arteries and veins. Vessels are dilated/enlarged with weakened vessel walls
that are prone to leakage and hemorrhage. Mutations in the KRAS gene in endothelial cells have been
identified in clinical AVM samples, leading to increased ERK1/2 phosphorylation and causing disruption of VE-
cadherin binding. Our goal is to create an AVM-on-a-chip model to better understand AVM pathology and to
uncover potential therapeutic drug targets.
Methods: KRASG12V ECs expressing mScarlet, normal GFP-expressing ECs, and fibroblasts were seeded in
hydrogel in the central chamber of a microfluidic platform. A ratio of KRASG12V ECs and normal ECs was used
to mimic the heterogeneity of AVMs and to create focal areas of mutated cells. Media flow was introduced into
the side channels connected to the tissue chamber by 1 port each. ECs self-assembled over the course of 7
days creating media-perfused microvessels. Vessel permeability/leakage was assessed via a fluorescent tracer.
Results: KRASG12V positive vessels showed significant larger diameter (47.3±28.5 μm) compared to
microvessels consisting of non-mutated ECs (18.2±7.2 μm). Interestingly, KRASG12V ECs accumulated in vessel
junctions (78.0±9.0%) – areas of known disturbed flow. Mimicking the characteristics of AVMs in vivo, vessels
containing the KRASG12V ECs were also more 2-fold more permeable to a fluorescent tracer than vessels
without the mutation by 30 minutes. This is likely due to the disruption of cell-cell VE-cadherin in KRASG12V
ECs, which will be quantified by immunofluorescence.
Conclusion and Significance: We were able to recreate in vitro the key characteristics of AVMs, namely vessel
enlargement and low barrier function, in a dynamic, perfused system. By developing an in vitro AVM in a
human relevant model, we can improve our understanding of the AVM pathophysiology and reliably assess
drug therapies.
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SYMPOSIUM
2021
Microfluidic arrays of dermal spheroids: a throughput screening

platform for skincare products
Zhengkun Chen, Sina Kheiri, Albert Gervorkian, Edmond W.K. Young, Valerie Andre, Ted Deisenroth, Eugenia
Kumacheva
Department of Chemistry, University of Toronto; Department of Mechanical and Industrial Engineering,

University of Toronto; BASF Beauty Care Solutions France S.A.S.; BASF Advanced Formulation Research North
America; Department of Chemical Engineering and Applied Chemistry, University of Toronto; Institute of
Biomaterials and Biomedical Engineering, University of Toronto
Rationale and Objectives: It is desirable to investigate alternative methods for evaluating cosmetics without
animal testing. Multicellular spheroids have emerged as a promising alternative in vitro model. Current
spheroids-based models have the limitations of a broad distribution of dimensions, lack of control on the
growth environments, and low screening capability. Here, we report a microfluidic (MF) skin-on-a-chip platform
for the growth of massive dermal fibroblasts spheroids (DFSs) and throughput screening of active ingredients
(AIs) of skincare products.
Methods: We utilized the self-digitization of a dense suspension of dermal fibroblasts to form a large array of
cell-laden droplets in the microwell MF device. These cell-laden droplets formed hundreds of uniformly sized
DFSs ready for screening within 48 hours. In a single chip, we accommodated 12 quadruplets devices, with
each quadruplet containing 200 microwells for the growth of 200 DFSs. For the case study, this DFSs-on-a-chip
model was used to screen the effect of vitamin C on the production of the two most important extracellular
matrix proteins, namely, collagen type I and fibronectin.
Results: By altering the size of the microwells and the loading densities of dermal fibroblasts suspension, we
achieve a 90% success rate in forming the cell-laden droplets and an 89% survival rate of the DFSs (without the
formation of a necrotic core). The computational fluid dynamic model demonstrated that the DFSs effectively
uptake, vitamin C, through advection and diffusion. We screened three doses of vitamin C and we spotted the
amount of Vitamin C that significantly enhances the secretion of collagen type I and fibronectin by the DFSs.
Conclusion and Significance: We demonstrated that our DFSs-on-a-chip platform offers a higher AI screening
capability and enables the screening of skincare products in a time-efficient manner, thus leading to a more
effective decision-making process. We envision that this platform can be further integrated into an industrial
setting for scaleup screening on AI as well as dermatological drugs.
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SYMPOSIUM
2021
A 3D vascularized liver model on the IFlowPlateTM*

Dawn S. Y. Lin, Boyang Zhang, and Shinichiro Ogawa
Department of Chemical Engineering, McMaster University; School of Biomedical Engineering, McMaster

University; McEwen Stem Cell Institute, University Health Network; Department of Laboratory Medicine and
Pathobiology, University of Toronto
Rationale and Objectives: The liver is a complex organ in our body that carries out numerous metabolic,
hormonal mechanisms. These mechanisms are complex and hard to recapitulate in vitro. In fact, many pre-
clinical models failed to predict the drugs’ metabolism in the liver. To accurately mimic liver functions for drug
development and study liver diseases, researchers will need a physiological model that has a functional unit
matching the natural hepatic lobule. Major building blocks for a unit like this should include hepatocytes, bile
ducts and hepatic sinusoids.
Methods: We customized a 384-well plate, also called IFlowPlateTM, which contains microchannels-connected
wells. Up to 128 3D tissues can be cultured using it. In this project, a liver cell line and hiPSC-derived
hepatocytes and cholangiocytes are used for building the parenchymal unit. An AggreWellTM800 plate is used
for liver spheroids formation. We also introduce endothelial cells (ECs) to build vascular networks that can span
over an area of 3.4x3.4 mm2 to support the parenchymal tissue. We use the ELISA, immunostaining, and
dextran perfusion techniques to check the level of albumin produced by liver cells cell phenotypes, bile
canaliculi formation, and perfusability and permeability of vascular networks.
Results: Our model supported liver cell growth in both single-cells and spheroids seeding conditions. The 3D
microenvironment in a hydrogel containing interstitial flow has been demonstrated to facilitate hiPSC-derived
liver cells’ maturation and differentiation by using the ELISA and immunostaining. Specifically, hiPSC-derived
cholangiocytes also migrated in the hydrogel to form duct-like aggregation. In addition, ECs formed perfusable
vascular networks which confine 70kDa dextrans, a comparable molecule to large proteins in the body, in the
luminal space with minimal leakage over time. In addition, the vascularized tissues can be physically extracted
from the well plate for implantation or downstream analysis.
Conclusion and Significance: We propose a 3D model consists of three integral parts of the liver: the
parenchymal tissue, bile ducts, and vascular networks. The presence of functional vascular networks allows us to
grow liver tissues with sufficient nutrients delivery and metabolic product exchange. We will run gene analysis,
cytokines secretion studies to further confirm cellular expressions.
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SYMPOSIUM
2021
HOnAChip (hypoxia on a chip): Insights into treatment efficacy*

Elena Refet-Mollof, Ouafa Najyb, Rodin Chermat, Audrey Glory, Julie Lafontaine, Phlip Wong, Thomas Gervais
Ecole Polytechnique de Montréal, Département de Génie Biomédical; Institut du Cancer de Montréal, Centre
de Recherche de l'Hôpital Universitaire de Montréal
Rationale and Objectives: Hypoxia is found in 50% to 60% of solid tumors and is a major contributor to
treatment resistance, which severely impacts patient survival. However, few drugs in development take hypoxia
into account, mainly due to the lack of good user-friendly translational models. In this study, we introduce a
simple microfluidic tool capable of reliably producing naturally hypoxic large tumor spheroids, and we
demonstrated how this model can be used to assess response to oxygen-dependent treatments such as TPZ
and RT.
Methods: We developed a non-perfused microfluidic chip allowing easy culture, maintenance, treatment, and
analysis of 240 sarcoma spheroids. 48h after cell seeding, spheroids were extracted for Western-Blots and
immunofluorescence assay to assess the expression and localization of hypoxic proteins Hypoxia Inducible
Factor 1 alpha and one of its downstream target, Carbonic Anhydrase IX. An in-silico model of oxygen
consumption was constructed using the IF results of CAIX expression in large spheroids and applied to small
spheroids. 48 h after cell seeding, spheroids were treated on-chip with two oxygen-dependent treatments, TPZ
and RT, as mono- and combined therapies.
Results: To date, we are still producing the largest spheroids on-chip (>750 µm of diameter), with a high
reproducibility. Our spheroids naturally express hypoxic proteins HIF1-a and CAIX.
Using IF results, our in silico model was able to predict absence of CAIX expression in smaller spheroids,
thereby confirming that spheroids must be of a certain size to naturally express sufficient levels of chronic
hypoxia. After 24h of treatment with TPZ, only the hypoxic regions showed a dose-response effect, confirming
the preferential cytotoxicity of TPZ to hypoxic cells. Combination therapy of TPZ and RT showed additive and
possible synergistic effects, which will be further investigated
Conclusion and Significance: Our device can produce homogeneous 240 jumbo naturally hypoxic spheroids.
Treatment with mono- and combined therapy of Tirapazamine and Radiotherapy on-chip can be easily
performed. Our tool can be used to assess efficacy of treatment modalities in presence of hypoxia, and may
serve as a predictive preclinical in vitro tool for in vivo studies.
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2021
High-throughput heart-on-a-chip platform for studies of SARS-

CoV-2-induced myocardial injury*
Qinghua Wu, Naimeh Rafatian, Jacob Smith, Karl Wagner, Sargol Okhovatian, Erika Wang, Yimu Zhao, Rick Lu,
Benjamin Fook Lun Lai, Teodor Veres, Milica Radisic
Institute of Biomedical Engineering, University of Toronto; Medical Devices, National Research Council Canada
Rationale and Objectives: We hypothesize that our new high-throughput heart-on-a-chip platform can serve as
an effective tool to create cardiac healthy and disease models for investigating the contractile dysfunctions
from SARS-CoV-2 infection and develop effective therapeutic strategies.
Methods: A novel 3D printing approach and new inks are developed to rapidly and automatically create the
heart-on-chip devices in a scalable manner. This platform enables us to investigate the pathogenesis
mechanisms of COVID 19 and identify specific cardiac cytopathic features. Direct myocardial infection and
multiple indirect factors will be performed and cardiac dysfunction will be measured in the platform, for
understanding the mechanisms triggering cardiac damage. Extracellular vesicles derived from human stem cells
will be investigated to assess whether they can impart cardioprotective effect on heart damage induced by
SARS-CoV-2.
Results: A heart-on-a-chip platform was developed as an excellent tool to study the contractile dysfunction of
human cardiomyocytes and mechanisms of the infection of SARS-CoV-2 into human cardiomyocytes. Biowires
was infected by SARS-CoV-2 to assess that cardiac tissues were directly targeted from SARS-CoV-2 infection.
The cardiac structure was identified myocardial damage by immunostaining. Dynamic contractile functions of
biowires were monitored daily after infection to observe the infection causes cytopathogenic effects on the
myocardium and brings new functional abnormities. Extracellular vesicles were investigated to have
cardiacprotective effect on the myocardium.
Conclusion and Significance: Our stem cell-based heart-on-a-chip models are probably the best around (at the
international scale) since we can very precisely quantify the contractile damage that occurs as a result of
infection and systematically study infection on different cell types that make up the heart and develop effective
therapeutic strategies, an experiment that is not possible in vivo.
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2021
Disease monitoring of protease activity using a reagentless

molecular pendulum*
Jenise B. Chen, Edward Tan, Jagotamoy Das, Sangeeta Bhatia, Shana O. Kelley
Department of Chemistry, University of Toronto; Massachusetts Institute of Technology
Rationale and Objectives: We are using an electrochemical molecular pendulum to determine the protease
activity for disease monitoring in vivo.
Methods: DNA barcodes injected into a mouse model are released into the urine by in vivo protease activity
and detected by the pendulum sensor. The molecular pendulum is a double-stranded DNA sequence with a
terminal redox reporter ferrocene. The kinetics are modulated through a positive potential bias, electrostatically
attracting the DNA to the surface and enabling ferrocene oxidation. Upon DNA barcode detection, the kinetics
are chronoamperometrically measured.
Results: We have developed a hybrid pendulum probe to enhance detection of nucleic acids. This hybrid probe
incorporates a region of flexibility, which enhances the modulation of the probe kinetics. Using this probe, we
show that it can detect DNA barcodes in the high nanomolar regime. This is detected chronoamperometrically
where kinetics can be measured and differentiated from the negative control.
Conclusion and Significance: A novel hybrid molecular pendulum that incorporates a flexible region has been
developed to enhance DNA detection. In the short term, we are working towards determining the limit of
detection. Ultimately, we are looking towards multiplexed detection of these barcodes in urine samples from
mouse models for infectious disease.
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SYMPOSIUM
2021
Automation of next generation sequencing sorkflows on a

centrifugal microfluidics platform*
Jimin Guo, Daniel Brassard, Caroline Miville-Godin, Maxence Mounier, Mojra Janta-Polczynski, Jason Ferreira,
Adrian Vesters, Ana Pilar, Nicholas Petronella, Nathalie Corneau, Denis Charlebois, Teodor Veres
Medical Devices Research Center, National Research Council Canada; Bureau of Microbial Hazards, Health
Canada; Canadian Space Agency
Rationale and Objectives: Genomic and transcriptomic sequencing is a maturing approach of microbiome
analysis, in vitro diagnosis, and public health surveillance. A bottleneck of its adaptation in settings with limited
resources lays in carrying out library preparation. NRC colleagues developed a centrifugal microfluidics liquid
handling platform, consisting of an instrument for transfering, mixing, and temperature control of liquids, and
thermoplastic cartridges for hosting chambers and microfluidics channels. Using this platform, we develop
automated workflows to prepare Illumina and Oxford Nanopore libraries.
Methods: We fabricated thermoplastic cartridges by injection molding. We programmed centrifugal force,

pneumatic force, and temperature control on the PowerBlade centrifugal microfluidics platform to sequentially
carry out liquid transfer, bead trapping and suspension, thermal incubation, and thermal cycling steps of various
library preparation procedures. We performed automated preparation of Illumina TruSeq libraries from a
defined mixture of three bacterial genomic DNA extracts. We sequenced the libraries on a NextSeq 550
system, alongside of a benchmarking library prepared manually. We performed a metagenomics analysis, as
well as a series of quality control assays of the libraries and of the reads.
Results: We observed ideal fragment sizes and comparable quality of the reads in both of the automated
libraries and the benchmarking library. At the chosen depth, we obtained full genomic coverage of all the input
bacterial species from each of the libraries. Reads in the two automated libraries constantly displayed less bias
towards GC-rich genomic islands, compared to the reads in the manual library. With a metagenomics analysis,
we recovered the identities of the three input species.
Conclusion and Significance: This development demonstrates the feasibility to automate complex liquid
handing and thermal cycling steps to carry out library preparation on the PowerBlade centrifugal microfluidics
platform. It serves as the basis of our series of efforts in implementing various genomics sequencing workflows
for space studies, food safety diagnostics, and COVID-19 pandemic surveillance.
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2021
Controlled shrinkage of microfluidically-generated microbubbles

by tuning lipid concentration*
Intesar Zalloum, Ali Paknahad, Raffi Karshafian, Michael C. Kolios, Scott S. H. Tsai
Department of Physics, Ryerson University; Department of Biomedical Engineering, Ryerson University; Keenan
Research Centre for Biomedical Science, St. Michael's Hospital; Institute for Biomedical Engineering, Science,
and Technology (iBEST) – A partnership between Ryerson University and St. Michael's Hospital
Rationale and Objectives: Microbubbles with diameters between O(1-5)𝜇m are used as contrast agents for
ultrasound imaging and drug delivery vehicles. Historically, microfluidically-generated bubbles are not used for
such applications because the size of the bubbles—which has a lower limit related to the microfluidic orifice
width—produced is too large (>10𝜇𝑚). Here, we report that microfluidically-generated bubbles can exhibit
differential shrinkage depending on the concentration of lipids. We study the effect of lipid concentration on
the amount of bubble shrinkage.
Methods: Encapsulated microbubbles are generated using a flow-focusing microfluidic device. The continuous
phase contains a mixture of lipids, glycerol, and Pluronic F-68 injected into the microfluidic device using a
syringe pump. A gas mixture containing octafluoropropane and nitrogen is injected in the disperse phase of the
microfluidic device using a pressure pump. The liquid flow rate and gas pressure are tuned until stable
microbubble formation of the desired size are achieved. We varied the concentration of the lipid solution from
0.9 mg/mL to 10 mg/mL.
Results: We demonstrate the synthesis of monodisperse bubbles with diameters of O(25−50) 𝜇𝑚. Once
microbubbles are generated, we posit that the bubbles become mechanically compressed to near-zero surface
tension as gas diffuses out due to Laplace overpressure within the bubble. Bubbles were found to dissolve to a
stable final diameter O(2-6) times smaller than their initial diameter, depending on the lipid concentration. This
is the first report for generating shrunk microbubbles in the sub-10 𝜇𝑚 range, by simply using more dilute lipid
solutions (<2 mg/mL) in the continuous phase.
Conclusion and Significance: In this work, we report for the first time, differential shrinkage of microbubbles by
varying the lipid concentration in the continuous phase. Future work will investigate the difference in acoustic
response and shell characteristics of bubbles that have undergone shrinkage compared to bubbles that have
not undergone shrinkage.
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2021
Rapid and automated detection of SARS-CoV-2 using isothermal

nucleic acid amplification*
Lidija Malic, Dillon Da Fonte, Christina Nassif, Maxence Mounier, Daniel Brassard, Andre Ponton, Matthias
Geissler, Matthew Shiu, Jason Ferreira, Emilie L. Gaudreau, Francois Normandin, and Teodor Veres
Life Sciences, Medical devices, Precision Diagnostics, National Research Council Canada
Rationale and Objectives: RT-qPCR performed in central laboratories is the gold standard for COVID-19
detection with sample-to answer time of 4-6 hours. The reverse transcription loop-mediated isothermal
amplification (RT-LAMP) is an emerging technology with potential to reduce testing time to under 1 hour. The
technique does not require specialized equipment and is thus more amenable to POC testing. We present a
rapid screening diagnostic test that could be completed in under one hour using an automated RT-LAMP assay
integrated with centrifugal microfluidic platform.
Methods: We have developed an assay, method and device architecture for automated sample-to-answer
detection of COVID-19 from swab patient samples. A protocol for on-chip viral RNA extraction based on
commercially available reagents for automated platform was demonstrated. In-house RT-LAMP Master Mix, as
well as primer sets targeting E and N genes were optimized enabling dry reagent formulation. A microfluidic
cartridge was designed for complete integration of RNA extraction and RT-LAMP assay using NRC’s patented
centrifugal microfluidic platform. The cartridge was fabricated in Zeonor using injection molding and validated
using both, spiked synthetic SARS-CoV-2 RNA, as well as real patient samples.
Results: SARS-CoV-2 RT-LAMP assay was developed and integrated with centrifugal microfluidic platform
allowing complete automation of sample-to-answer workflow that can yield diagnostic results in under one
hour. The assay included 10 min sample preparation (lysis and RNA extraction), followed by 30 min RT-LAMP
reaction and colorimetric detection. The assay was validated using synthetic SARS-CoV-2 RNA and was capable
of detecting low copy numbers typically seen in patients at the onset of symptoms (LOD of 0.5 copies per μl)
while having a large dynamic range (102-109 copies per mL). Further assay validation using 3 positive and 3
negative patient samples demonstrated the platform specificity and sensitivity.
Conclusion and Significance: An automated COVID-19 diagnostic test was developed allowing highly sensitive
detection of SARS-CoV-2 RNA (LOD < 1 copy per μl) within 40 minutes. Given the high sensitivity and
specificity of the test, the platform could be deployed in settings where repeat testing and/or rapid turnaround
times are important, such as outbreak control or remote communities.
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2021
Molecular assay for instrument-less SARS-CoV-2 rapid diagnostic

for saliva*
Warren C. W. Chan
Institute of Biomedical Engineering, University of Toronto
Rationale and Objectives: Successfully diagnosing SARS-CoV-2 is critical to containing the spread of COVID-
19. The most sensitive approach is typically a genetic assay, which requires multiple steps to isolate, extract,
amplify, and detect the virus. Simplifying these steps will increase the efficacy of diagnosing SARS-CoV-2 and
lead to greater prevention of spread.
Methods: We propose to optimize and perform a proof-of-concept study that combines microfluidic and
molecular assays to detect SARS-CoV-2 in saliva by a colour change within 30 minutes at a minimum sensitivity
of 10-18 mol. The microfluidic system contains compartments with different steps of the molecular assay. This
assay consists of reagents that will amplify the SARS-CoV-2 genetic material (e.g. E gene) by 10 thousand
fold. The amplified genetic material will bind to 60 nm gold nanoparticles coated with single stranded DNA
(ssDNA) that is complementary to SARS-CoV-2 gene. The colour of the solution will change because the gold
nanoparticles will assemble onto the amplified SARS-CoV-2 E gene. The clustering of the gold nanoparticles
changes how their electrons are moving, leading to different absorbance properties. This physical process
results in a colour change in the gold nanoparticle solution from red to blue. A positive detection occurs when
the solution is blue. We propose to develop a microfluidic chip where different chambers control each of the
diagnostic assay reactions. We will load the assay reagents (which consist of enzymes, salts, and gold
nanoparticle-conjugates) in each chamber. Individual reactions will occur by flowing the patient sample from
one chamber to the next in a disposable device that can be actuated with the use of an instrument in the
simplest implementation.
Results: We have successfully assessed each step of the molecular assay. In particular, we have optimized the
chemistry of labeling gold nanoparticles with SARS-CoV-2-capturing ssDNA and have characterized their
performance with synthetic genetic targets. We are starting to assess clinical samples using this gold
nanoparticle probe system after extraction, transcription, and amplification.
Conclusion and Significance: Developing diagnostic technologies that can be accessible to the masses is critical
to preventing the spread of SARS-CoV-2. Our assay performs well. We are starting to design the microfluidic
chips and to incorporate our assay system into the chip.
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2021
Latex agglutination tests for rapid, instrument-free COVID-19

diagnostics in saliva*
Alexandros A. Sklavounos, Joshua Dahmer, Mohammed A. A. Abdullah, Anthony Yong, Sidharth Gupta, Jose
Gilberto Camacho Valenzuela, Martin A. Rossotti, Keith Morton, Matthew Shiu, Alex Mariakakis, Jamshid Tanha,
Jean Labrecque, Teodor Veres, Aaron R. Wheeler
Dept. of Chemistry, University of Toronto; Donnelly Centre for Cellular and Biomolecular Research, University of
Toronto; Dept. of Computer Science, University of Toronto; Inst. of Biomedical Engineering, University of
Toronto; Human Health Therapeutics Research Centre, National Research Council Canada; Life Sciences
Division, National Research Council of Canada; Dept. of Biochemistry, Microbiology & Immunology, University
of Ottawa
Rationale and Objectives: The COVID-19 pandemic has created a need for rapid and ubiquitous testing. Latex
agglutination tests (LATs) are a simple, and instrument-free methods for detecting infectious agents. LATs are
routinely used for detecting targets from bacterial antigens to viruses. Therefore we chose to develop an LAT
assay for detecting the SARS-CoV-2 virus, which would enable monitoring of active infection cases. While the
virus is primary sampled with a nasopharyngeal swab, in an effort to improve user experience, we adapted our
assay to be used in saliva.
Methods: LATs rely on a process called agglutination, which is the aggregation of latex particles in presence of
an analyte. The result of the assay can be distinguished by eye or can be digitized and processed using a digital
camera and image analysis. Latex particles, in a range of particle sizes, were modified with antibodies that
identify the SARS-CoV-2 antigens using several conjugation methods. A large sample set of antibodies against
S1, S2 and N protein epitopes of SARS-CoV-2 were evaluated. Suspensions of sensitized particles were mixed
with solutions containing inactivated virus and the progress of agglutination was recorded over time using an
in-house built imaging unit.
Results: A portable imaging unit for 96-well well-plates was developed and images were processed for
automated evaluation of the agglutination result. Using machine learning algorithms, the degree of
agglutination was also defined and used to evaluate and optimize the parameters of our assay (bead size, bead
color, conjugation method, particle loading, particle density, antibody epitope, buffer, ionic strength). The top
candidates were selected for the assays in saliva. The saliva dilution was optimized and additives such as salts
and surfactants were used to maintain the performance of our assay with the new matrix.
Conclusion and Significance: An LAT assay was developed for the rapid and instrument-free detection of SARS-
CoV-2 in saliva. This is an important milestone towards providing easy and reliable detection of the virus to the
general public. As a next step we will be challenging our system with patient samples in collaboration with Sick
Kids Hospital. The ultimate goal is to feature our assay in a chewable lollipop-like device for non-invasive
diagnostic of COVID-19 in saliva.
Scheduled for: August 27, 2021 from 1:30-1:45 *Competing in CRAFT People’s Choice Award
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2021
An automated microfluidics glucose meter interface for point-of-

care gene circuit-based diagnostics*
Evan Amalfitano, Jennifer Doucet, Moiz Charania, Kebin Li, Matthew Shiu, Javier Hernandez-Castro, Abdul
Wasey, Francois Normandin, Caroline Miville-Godin, Teodor Veres, Keith Pardee
Leslie Dan Faculty of Pharmacy, University of Toronto; National Research Council Canada; Centre for Research
and Applications in Fluidic Technologies, University of Toronto; Department of Mechanical and Industrial
Rationale and Objectives: As the COVID-19 pandemic has shown, new disease outbreaks require rapid
diagnostics as part of a coordinated response. Centralized conventional diagnostics, while effective, often have
cost, time, and equipment limitations that can reduce their usefulness, particularly during sudden demand
surges and in remote areas. To address these issues, we are developing a new low-cost automated microfluidic
diagnostic system that provides an output in the form of glucose, allowing the test results to be measured with
an off-the-shelf glucose meter.
Methods: Gene circuits (molecular switches activated by a pathogen) have been designed for detection of
SARS-CoV-2, generating a glucose-producing reporter enzyme in response. These gene circuits are combined
with a cell-free protein expression system, containing the necessities for transcription and translation. In
conjunction with sample preparation reagents and an isothermal amplification reaction, this allows detection of
SARS-CoV-2 from saliva samples. The resulting glucose produced by the reporter enzyme is then quantified on
a standard glucose meter to read out the test results. All components of this system will also be integrated into
a low-cost, portable, and automated microfluidic device.
Results: Gene circuits for the detection of SARS-CoV-2 have been developed and validated using a glucose-
producing reporter enzyme. Sample preparation and isothermal amplification elements have also been
validated using heat-inactivated SARS-CoV-2 virus. Significant progress has also been made in developing the
microfluidic device, including new systems for metering and mixing liquids developed by the microfluidics team
at the NRC facility in Boucherville, QC. Focus on cost reduction has resulted in an estimated per test cost of
less than $20. Challenges have arisen in acquiring clinical samples for testing, but these are being addressed.
Conclusion and Significance: The overall system workflow has been validated using contrived samples on the
benchtop. Next steps include testing the system on the microfluidic device and with real patient samples. Work
is also underway to adapt the platform for the detection of specific SARS-CoV-2 variants. The system is
programmable and can be easily adapted to meet future diagnostic needs.
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ABSTRACTS
2021
POSTERS 33
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2021
Poster #1: Integration of microfluidic spheroid-on-a-chip platform

with machine learning
Ilya Yakavets, Sina Kheri, Matteo Aldeghi, Jennifer Cruickshank, Jose Dario Perea Ospina, Alán Aspuru-Guzik,
David Cescon, Eugenia Kumacheva
Department of Chemistry, University of Toronto; Department of Mechanical & Industrial Engineering, University
of Toronto; Chemical Physics Theory Group, Department of Chemistry, University of Toronto; Department of
Computer Science, University of Toronto; Vector Institute for Artificial Intelligence; Princess Margaret Cancer
Centre, University Health Network; Institute of Biomedical Engineering, University of Toronto; Department of
Chemical Engineering and Applied Chemistry, University of Toronto
Rationale and Objectives: Cancer remains a devastating disease in North America and worldwide. Combination
chemotherapies offer the potential for the improvement of therapeutic responses of cancer patients by utilizing
multidrug cancer treatment. Spheroids, sub-millimeter clusters of cancer cells, recapitulating many properties of
tumors, show great promise in the development of multidrug therapies. Here, we report a development of a
physiologically relevant spheroid-on-a-chip microfluidic platform to identify clinically valuable multidrug
regimens for cancer treatment.
Methods: Breast cancer spheroids were grown in tissue-mimetic hydrogels using breast carcinoma MCF-7 cells.
Spheroids grew under the physiological flow for 48 hours and then were subjected to individual drugs and their
combinations by continuous perfusion for 72 hours. The response of spheroids to drugs was evaluated by
live/dead assay using fluorescence microscopy. To make a decision about the best drug combination in a time-
effective manner, the spheroid’s response was fed into a machine learning algorithm (Phoenics), a recently
developed Bayesian neural network optimizer. Based on the spheroid's response, the algorithm adaptively
learned to propose drugs with higher and higher anticancer activity.
Results: Microfluidic device accommodated 200 spheroids in the100 μm microwells that are organized in 4
parallel rows, each comprising 50 cylinder-shaped microwells. The spheroids were formed from MCF-7 cells in a
fibrillar hydrogel matrix composed of gelatin (providing bioadhesive ligands) and cellulose nanocrystals. After
48 hours of culture, spheroids show high viability (live/dead assay) and display established E-cadherin cell-cell
junctions between the cells. We estimated dose-response curves for doxorubicin and paclitaxel after 72 hours
of perfusion. The dose-response profiles for drugs were then fed into a Phoenics, which proposed conditions
for combinational treatment.
Conclusion and Significance: We developed an innovative microfluidic cancer organoid-on-a-chip platform

utilizing a biomimetic hydrogel matrix, uniformly sized spheroids, and spheroid growth and drug delivery under
close-to-physiological flow. Integration of machine learning will provide time-effective discovery and
optimization of multidrug therapies for cancer treatment.
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2021
Poster #2: Optimizing tumor-macrophage co-culture on a 3D

tumor platform
Ziting (Judy) Xia, Ileana Co, Alison McGuigan
Rationale and Objectives: The interactions between tumor-associated macrophages (TAM) and pancreatic
ductal adenocarcinoma (PDAC) are the leading cause of PDAC's poor prognosis. To study such interactions, the
McGuigan lab has fabricated a 3D tumor model, TRACER, which can be used as a platform for the PDAC-TAM
co-culture. This current study aims i) to develop a co-culture media that does not polarize macrophages
prematurely and ii) to optimize digestion protocols to enable recovery of viable cells from the TRACER scaffold
for downstream analysis.
Methods: PDAC organoids and TAMs were separately cultured before mixing together in TRACER scaffolds.
Because the transcriptome of TAMs is more sensitive, pure macrophage media RPMI was used for the co-
cultures. The monoculture PDAC media was modified to allow PDAC cells to adapt to the new co-culture
media. Five mixing ratios (0%, 25%, 50%, 75%, 100% RPMI) were tested to identify the one leading to the
highest tumor maturation indicated by CK19 expression. For digestion, the type of digestive enzymes
(collagenase IV, XI, and Liberase), concentration, incubation time and mechanical shaking were explored in
various combination. Fluorescence emission of Calcein/PI staining was used as metric for viability and recovery.
Results: To examine the media conditions, CK19 expression was quantified for PDAC organoids upon co-
culture media switch. Fluorescent confocal imaging revealed that PDAC organoids cultured in 25% and 75%
RPMI mixing ratios showed a continuously increasing trend of CK19 expressions when in RPMI, indicating a
successful adaptation to the new media over time. For the digestion protocol, Calcein/PI emission indicated
Liberase and Collagenase IV as the optimal digestive enzyme at 1000 µg/mL and 300 U/mL respectively.
Further, at an incubation time of 45 min and shaking of 300 RPM, the recovered cells achieved 90% viability.
Conclusion and Significance: The optimization of the co-culture media and digestion protocol is significant in
aiding the successful fabrication of TRACER. It also demonstrates TRACER's potential as a de novo 3D in vitro
platform capable of facilitating complex tumor–immune cell–cell interaction queries while recapitulating the in
vivo tumor biology, such as hypoxic and metabolic gradients.
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2021
Poster #3: A 3D engineered tumour-tissue model for image-

based measurements
Nila C Wu, Jose L Cadavid, Simon Latour, Xinzhu Tan, Alison P McGuigan
Institute of Biomedical Engineering, University of Toronto; Chemical Engineering and Applied Chemistry,
Rationale and Objectives: Cancer is ranked as a leading cause of death worldwide. However, failure rates for
cancer drug discovery are significantly higher than in other therapeutic areas. This can be attributed to the use
of non-representative pre-clinical cancer models and assays to evaluate compound efficacy. This includes using
non-patient samples and whole-well, mono-culture, end-point assays. Here, we address these limitations by
developing a disease-relevant 3D engineered tumour-tissue platform in a 96-well plate format, suitable for
image-based analysis.
Methods: We have scaled-up our previously published 3D cell culture GLAnCE platform, to a 96-well plate
version to increase sample throughput. Fluorescently-labeled tumour cells and tumour stromal cells are seeded
into 96-GLAnCE in mono-culture or co-culture conditions, and growth of each cell population is tracked and
quantified over-time using a high-content widefield microscope and automated image analysis algorithms. To
assess response to therapy, 96-GLAnCE cultures are treated with chemotherapy drug, and subsequent
regrowth is monitored within a drug-treated microenvironment. Due to its miniaturization, each tumour-tissue
requires low sample volumes, thereby enabling the use of patient-derived organoids (PDOs).
Results: As a result of 96-GLAnCE’s unique seeding method, uniform and thin layers of low volumes of hydrogel
are generated, that outperform gel domes and plugs for consistent image analysis. To demonstrate the
platform’s suitability with image-based quantification, we show multi-cell engineered tumour-tissues can be
generated and cell growth of each population can be robustly monitored over-time. Next, we establish the
regrowth assay in 96-GLAnCE, where we track cancer cell signal intensity over-time after treatment with
different concentrations of chemotherapy drugs. Moreover, we incorporated GFP-expressing PDOs into 96-
GLAnCE, and demonstrate co-culture growth and PDO regrowth after chemotherapy treatment.
Conclusion and Significance: 96-GLAnCE facilitates continuous, image-based measurements of cell growth in
multi-cell 3D engineered tumour-tissues and regrowth after treatment with chemotherapy drugs, in a 96-well
platform. Using our technology would enable compound screening in a disease-relevant cancer model using
patient-derived samples, towards successfully identifying a novel cancer drug.
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2021
Poster #4: Design of islet-on-a-chip to measure insulin, glucagon

and GLP-1
Yufeng Wang, Romario Regeenes, Amir G. Moghadam, Jonathan V. Rocheleau
Institute of Biomedical Engineering, University of Toronto; Department of Physiology, University of Toronto;

Toronto General Research Institute, University Health Network
Rationale and Objectives: A complex network exists within pancreatic islets and regulates hormone secretion.
This intra-islet signalling is crucial in maintaining glucose homeostasis and is impaired in type 2 diabetes.
Investigating dynamic islet secretion will help tease apart the underlying regulatory mechanisms. This project
aims to design an islet-on-a-chip to simultaneously measure multiple hormone secretion from individual islets
with high temporal resolution to investigate the interplay between different islet cells, and their responses to
chronic hyperglycemia.
Methods: C-peptide, glucagon and GLP-1 secretion will be simultaneously monitored using fluorescence-
anisotropy of a competition assay that uses a corresponding synthetic peptide conjugated to a fluorescent. An
islet-on-a-chip will be designed to hold islets stable during imaging while providing sufficient residence time for
the on-chip assay. To validate the design, we will test it against standard off-chip bulk assays, and correlate c-
peptide, glucagon and GLP-1 responses to glucose of healthy human islets. To evaluate hormone secretion
under metabolically stresses, we will investigate islet secretion on-chip from human islets treated with high
concentrations of glucose, or other metabolites for at least 24 hours.
Results: We have developed the immunoassay for c-peptide with the proper probe design and maximized
sensitivity. The kinetics of the assay has also been characterized and incorporated into the microfluidics design.
With the aid of modeling in COMSOL Multiphysics, we are in the process of finalizing. The same workflow will
be applied for developing the on-chip assay for glucagon and GLP-1. Currently, the major challenge lies in
designing the proper post-imaging analysis method to perform non-equilibrium measurement of the assay.
Conclusion and Significance: The general protocol developed for c-peptide assay will significantly accelerate
the process for both glucagon and GLP-1. With the on-chip hormone detection, both the measurement speed
and temporal resolution will be enhanced when investigating dynamic islet secretion. Ultimately, the islet-on-a-
chip will provide a powerful research tool in the islet biology field.
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2021
Poster #5: A high-content compound screening model for

progressive cardiomyopathy
Erika Yan Wang, Uros Kuzmanov, Jacob B. Smith, Wenkun Dou, Naimeh Rafatian, Benjamin Lai, Rick Lu,
Qinghua Wu, Joshua Yazbeck, Yu Sun, Anthony Gramolini, and Milica Radisic
Institute of Biomedical Engineering, University of Toronto; Translational Biology and Engineering Program, Ted
Rogers Centre for Heart Research; Department of Chemical Engineering and Applied Chemistry, University of
Toronto; Toronto General Hospital Research Institute, University Health Network; Department of Mechanical
and Industrial Engineering, University of Toronto
Rationale and Objectives: Angiotensin II (Ang II) presents a critical mediator in various pathological conditions
such as non-genetic cardiomyopathy. Osmotic pump infusion in rodents is a commonly used approach to
model cardiomyopathy associated with Ang II. However, profound differences in electrophysiology and
pharmacokinetics between rodent and human cardiomyocytes may limit predictability of animal-based
experiments. This study investigates the application of an Organ-on-a-chip (OOC) system in modeling Ang II-
induced progressive cardiomyopathy.
Methods: The disease model is constructed to recapitulate myocardial response to Ang II in a temporal
manner. Along with mapping of cytokine secretion and proteomic profiles, this model presents an opportunity
to quantitatively measure the dynamic pathological changes that could not be otherwise identified in animals.
Further, we present this model as a testbed to evaluate compounds that target Ang II-induced cardiac
remodeling.
Results: The long-term tissue cultivation and non-invasive functional readouts enable monitoring of both acute
and chronic cardiac responses to Ang II stimulation. Through assessing the effects of losartan, relaxin, and
saracatinib, the drug screening data implicated multifaceted cardioprotective effects of relaxin in restoring
contractile function and reducing fibrotic
remodeling.
Conclusion and Significance: Overall, this study provides a controllable platform where cardiac activities can be
explicitly observed and tested over the pathological process. The facile and high-content screening can
facilitate the evaluation of potential drug candidates in the pre-clinical stage.
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2021
Poster #6: Targeting highly potent therapeutic T cells by

immunomagnetic cell sorting
Zongjie Wang, and Shana O. Kelley
Institute of Biomedical Engineering, University of Toronto; Latner Thoracic Surgery Research Laboratories, The
Sprott Department of Surgery, University Health Network; Department of Pharmaceutical Sciences, University of
Toronto; Department of Biochemistry, University of Toronto; Department of Chemistry, University of Toronto
Rationale and Objectives: Tumor-infiltrating lymphocytes (TILs) are a subset of white blood cells that have left
the bloodstream and migrated toward a tumor. These cells – when isolated from tumors and expanded – have
significant antitumor activity and TILs have been widely used as source cells for adoptive cell therapy (ACT). The
success of ACT requires the recovery and selection of highly potent TILs. However, TILs are rare in tumors and
difficult to be isolated efficiently with existing protocols, leading to limited characterization and optimization of
their potency.
Methods: We hereby describe a highly tunable microfluidic platform, termed microfluidic affinity targeting of
infiltrating cells (MATIC), for the recovery and expansion of specific TIL subpopulations from tumor tissue based
on quantitative immunomagnetic sorting. MATIC uses a series of modular microfluidic chips fabricated by
scaled 3D printing in a cleanroom-free fashion and designed for configurable, quantitative, and volumetric cell
separation. We describe the application of this approach for the characterization of the potency of TILs based
on the expression of CD8 and an immune checkpoint (referred to as target A below) through in vitro molecular
assay and in vivo adoptive transfer models.
Results: We were able to achieve a yield rate of 40 microfluidic chips at the cost of $10 per chip to meet the
demand of scaled applications. In studies comparing the properties of CD8+ T cells isolated from tumors, we
show that the MATIC platform offers higher yields (up to 30-fold) and levels of potency (up to 30%
improvement in median survival) than other marker-specific approaches. We then pursued the targeting of TILs
based on target A - we identified that moderate expression of target A defines a unique population of TILs that
is stem-like, self-renewable, and tumor-specific. This subpopulation of TILs is highly potent and controls tumor
growth in vivo (up to 60% improvement in median survival).
Conclusion and Significance: Our studies highlight the importance of cell isolation methods in the
establishment and manufacturing of cell therapy products. The improved recovery from MATIC is beneficial for
the efficacy and translation of adoptive cell therapy. We also identified a unique biological signature to identify
highly potent TIL subpopulations.
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2021
Poster #7: In vitro electrode sensors on porous membrane for

biological barriers
Alisa Ugodnikov, Oleg Chebotarev, Henrik Persson, Craig A. Simmons
Rogers Centre for Heart Research
Rationale and Objectives: Traditional methods to assess endothelial barriers – permeability tracer assays,
transendothelial electrical resistance (TEER) – require invasive handling and cannot directly measure monolayer
integrity in co-culture setups typical to organ-on-chip systems. Conversely, electrical cell-substrate impedance
sensing (ECIS) measures impedance of cells grown on electrodes. Here we report integration of ECIS
electrodes on porous membrane, enabling non-invasive, direct monitoring of endothelial cells in co-culture
setup, and validation against TEER.
Methods: Gold electrodes were fabricated on porous membrane inserts using hot embossing and standard UV
lithography techniques. Membrane electrodes were coated with fibronectin (25 μg/cm2) to promote cell
adhesion for primary human umbilical vein endothelial cells (HUVECs). A lock-in amplifier (SR850, Sanford
Research Systems) was used to both generate alternating current (62.5 – 64,000 Hz) and measure impedances.
Results: Frequency scans of confluent HUVEC monolayers on ECIS membrane inserts showed normalized
resistance values peaking at 4 kHz, consistent with accepted measurement frequency for HUVECs in
conventional glass substrate setups. ECIS resistance (4 kHz) for HBMEC monolayers correlated with FITC-
dextran permeability measurements, validating the ECIS membrane method. Sensitivity was assessed by
monitoring HUVEC growth on working electrodes of varying sizes (d = 250, 500, 750 μm), as well as response of
confluent monolayers to 5 mM EGTA, with greatest sensitivity to changes in barrier integrity (4 kHz) observed
using the smallest electrodes. ECIS measurements were correlated to TEER measurements taken in same
devices.
Conclusion and Significance: Our platform produces resistance outputs and electrode size-dependent
impedance trends consistent with conventional glass substrate ECIS setups, and allows for simultaneous
validation against gold standard techniques used to evaluate co-culture systems. This method is compatible
with integration into a microfluidic platform for organ-on-chip culture.
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2021
Poster #8: New membrane-based, water-in-water microdroplet

production platform
Shyan Thompson, Dae Kun Hwang
Department of Chemical Engineering, Ryerson University; Keenan Research Centre for Biomedical Science, St.
Michael’s Hospital; Institute for Biomedical Engineering, Science and Technology (iBEST),
A Partnership Between Ryerson University and St. Michael’s Hospital
Rationale and Objectives: Aqueous two-phase system (ATPS) microdroplets have received great attention due
to their biocompatibility, which is highly desirable for biomedicine and biological applications. However, their
low interfacial tension restricts conventional production mechanisms. Recently developed passive, hydrostatic
pressure-based methods are preferred over active approaches using external forces, although they are difficult
to scale-up. Thus, we developed a new passive membrane-based ATPS platform that is scalable to produce
monodispersed droplets.
Methods: The microfluidic chip is comprised of three components: top channel, bottom channel and
membrane with micropores. Standard soft lithography was used for the top (60-µm tall, single-step PDMS mold)
and bottom channel (120-µm tall, two-step PDMS mold equipped with membrane seating and supports). Slit
channel lithography, developed in our group, was used to fabricate a membrane with an array (3 x 1) of
micropores. The membrane was inserted into the bottom channel and dried, before air plasma treatment and
PDMS-PDMS bonding. Syringe pumps were used to introduce the dextran (DEX) and polyethylene glycol (PEG)
phases of the ATPS into the membrane-integrated microfluidic channel.
Results: We successfully produced microdroplets by adjusting the flowrates of discontinuous DEX and
continuous PEG. The DEX phase flowed from top channel to membrane pore to bottom channel. Upon
entering the PEG phase in the bottom channel, the co-flow streams produced DEX droplets. Alongside the
droplet regime, ripple, thread and backflow were observed, and changing the membrane pore size allowed for
distinguishable differences in regime and transition patterns based on flowrate. The system was capable of
producing highly monodispersed 17 to 180 µm-droplets with a coefficient of variation (CV) as low as 2.6%.
Different pore shapes with identical characteristic lengths were utilized and found to act alike.
Conclusion and Significance: Our ATPS, membrane-based droplet production platform enables the production
of biocompatible monodispersed DEX phase droplets. The use of microporous membranes and syringe pumps
allow for accessible scale-up. Larger arrays of pores working in parallel can be readily considered to
compensate for the low throughout.
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2021
Poster #9: Microfluidic disease models of the blood-brain barrier

Lily Takeuchi, Craig A. Simmons
Institute of Biomedical Engineering, University of Toronto; Department of Mechanical and Industrial

Engineering, University of Toronto; Translational Biology and Engineering Program, Ted Rogers Centre for
Heart Research
Rationale and Objectives: The blood-brain barrier (BBB), a semi-permeable barrier of specialized endothelial
cells, plays a role in regulating entry of molecules to the brain. The BBB presents a challenge to the delivery of
therapeutics, and its function has been demonstrated to be altered by shear flow and under disease states.
There is an unmet need for models that recapitulate the pathological barrier and accurately predict drug
bioavailability and efficacy. Here, we aim to develop stem cell-derived models of the blood-brain barrier for
drug screening applications.
Methods: Immortalized human brain endothelial cells (BECs) were characterized for BBB phenotype to serve as
a benchmark for future models involving induced pluripotent stem cell-derived BECs from patients with
Alzheimer’s Disease and hereditary stroke. Key characteristics of the BBB such as barrier strength and
expression of junctional proteins were assessed by transendothelial electrical resistance and
immunofluorescence, respectively. Permeability of the barrier to small and large molecules was assessed using
fluorescence tracer assays and function of specific influx and efflux transporters were assessed using substrate
uptake and inhibition assays.
Results: Immortalized BECs achieved maximum barrier strength after 5 days and yielded transendothelial
electrical resistance values of 159 ± 45 Ω*cm2. The cells yielded permeability coefficients within range of
literature-reported values of 1.5 ± 0.64 x 10-7 cm/s and 8.4 ± 0.18 x 10-8 cm/s for doxorubicin and 4 kDa FITC-
dextran, respectively. The cells expressed junctional proteins ZO-1, VE-cadherin, claudin-5, and occludin at cell-
cell junctions, demonstrated alignment in the direction of shear flow in a microfluidic platform, and presented
functional p-glycoprotein efflux transporter function and receptor mediated transcytosis of low density
lipoprotein.
Conclusion and Significance: Immortalized BECs present the unique features of the BBB endothelium and
provide a suitable cell source for modelling the healthy BBB. This work will be important towards providing a
benchmark against BECs from patients with neurological disorders and advancing our understanding on how
alterations to BBB phenotype due to disease may impact efficacy of therapeutics.
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2021
Poster #10: Fast and bubble free PCR on centrifugal microfluidics

Tae-Hyeong Kim, Dillon Da Fonte, Christina Nassif, Daniel Brassard, Teodor Veres
Life Sciences, Medical Devices, National Research Council Canada
Rationale and Objectives: PCR has been the gold standard for the detection of infectious diseases. However, it
requires bulky instrument with trained personnel and such drawbacks have limited the application of PCR in
low-resource settings. Microfluidic PCR has been offering a unique advantage, the probability for automation to
minimize the need for resources, but the bubble trapping issue has been the critical challenge. Here we report
a fast and bubble-free PCR microfluidic device to overcome conventional drawbacks.
Methods: The microfluidic design was milled with a CNC milling machine and laminated using double-sided
adhesive tape. On a microfluidic layout, a radially long reaction chamber was designed and outward end was
connected to the pressure port for the pneumatic control. Two oils of different densities (mineral oil and
fluorinated oil) were utilized to induce the density-based separation. PCR mixture and both oils were added to
the reaction chamber and sealed. A device was installed on PowerBlade capable of spinning a device, applying
active pneumatic pressure, and heating. Then, PowerBlade was operated by programmed protocol to run an
assay automatically.
Results: In the reaction chamber, two oils and PCR mixture were separated by the density during rotation.
Mineral oil moved inward while heavier fluorinated oil moved outward. PCR mixture of medium density was
positioned between two oil layers removing bubbles due to centrifugal force. A mineral oil layer could prevent
evaporation and a fluorinated oil assisted radial movement of PCR mixture in the reaction chamber. Positive
pneumatic pressure was applied through the pressure port and it pushes a fluorinated oil. It leads to movement
of PCR mixture’s radial position and pressure was controlled to reciprocate between two heating zones for PCR.
Conclusion and Significance: We established a fast and bubble-free microfluidic PCR system on PowerBlade.
Centrifugal force aided the bubble removal effectively while active pneumatic pressure control allowed easy
radial movement of PCR mixture to achieve space domain PCR. This technique would be a useful tool to
employ PCR in low-resource settings.
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2021
Poster #11: Vibration in preventing breast cancer bone

metastasis
Xin Song, Chun-Yu Lin, Yaji Ke, Liyun Wang, Lidan You
Department of Mechanical and Industrial Engineering, University of Toronto; Institute of Biomedical

Engineering, University of Toronto; Department of Chemical Engineering, University of Toronto; Department of
Mechanical Engineering, University of Delaware
Rationale and Objectives: Bone metastasis occurs in 75% of patients with advanced breast cancer. Metastasized
cancer cells alter bone remodelling, leading to incurable bone lesions. Exercise as a common cancer
intervention is often physically challenging for the elderly (>65yrs). Low magnitude (<0.3g) high frequency
(>30Hz) (LMHF) vibration, which has been shown to promote bone health, could serve as a safer alternative.
This project aims to study the role of osteocytes, the mechanosensors in bone, in regulating breast cancer
migration under LMHF vibration.
Methods: We developed a microfluidic co-culture platform which allows real-time cell-cell crosstalk between
different cell populations. Metastatic breast cancer MDA-MB-231 cells were cultured in the lumen channel lined
with endothelial cells (HUVEC), which was adjacent to the channel seeded with osteocyte-like MLO-Y4 cells.
LMHF vibration was set at 0.3g and 60Hz for 1 hour, which has been previously shown to have promising
effects in reducing osteoclast formation. Cells in the platform received vibration 1 hour per day for 3 days. The
migration rate of breast cancer was monitored.
Results: Our previous monoculture results showed that vibration on breast cancer cells only does not change
their migration. However, when co-culturing breast cancer cells with endothelial cells and osteocytes in the
microfluidic platform, cancer migration was reduced by 40% under vibration.
Conclusion and Significance: LMHF vibration activated osteocytes, leading to cellular crosstalk that reduces
breast cancer migration. The microfluidic platform provides insights into the impact of vibration on bone
metastasis. This research seeks to unleash the benefits of vibration for elderly breast cancer patients, leading to
a safe treatment for the high-risk population.
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2021
Poster #12: Increasing automation in the cardiac biowire II

platform
Jacob Smith, Milica Radisic
Department of Chemical Engineering & Applied Chemistry, Institute of Biomedical Engineering, University of
Toronto
Rationale and Objectives: The Organ-on-a-Chip Biowire II platform has been validated to be a robust model of
both healthy and diseased cardiac tissue, and enables advanced drug testing methodologies in a 3D cardiac
tissue. Functional 3D cardiac tissue readouts from the platform include contraction force, calcium transients,
electrical properties. The present analysis pipeline includes a variety of manual processing steps. These steps
include: brightfield tissue dimensional measurement, wire deflection calculation, calcium transient detection
and data processing. As the Biowire II platform continues to increase in scalability with greater numbers of 3D
tissues, there is a need to simultaneously improve the throughput of the analysis pipeline.
Methods: To enhance tissue measurement speed, an automated segmentation program was developed to
detect and extract the tissue images from video recordings. This was accomplished using a machine learning
algorithm that was trained on manually segmented images generated by expert users. The algorithm was
created using the ImageJ image analysis software developed by the National Institute of Health, alongside
MATLAB by MathWorks.
Results: Using this system, dimensional measurements can be rapidly extracted from Biowire II tissue images,
eliminating the need for a user to manually measure potentially hundreds of images in an experiment. The
current focus of the project is adjusting the segmentation system to extract wire deflection data and to develop
a calcium transient detection method to accurately select true calcium transients. Once established, the entire
Biowire II pipeline will be collapsed into a simple streamline system where the only user action required is
sample selection before receiving a complete data output file.
Conclusion and Significance: These changes will significantly improve the throughput of the Biowire II platform,
by eliminating the time-consuming manual processing steps of the analysis pipeline. This will greatly increase
the quantity of samples that can be run, enabling more compounds to be rapidly tested which increases the
discovery potential for new therapies.
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Poster #13: UV-induced polymerization to synthesize worm-like

particles
Sinthuran Jegatheeswaran, Dae Kun Hwang, Mauricio Terebiznik
Department of Chemical Engineering, Ryerson University; Keenan Research Centre for Biomedical Science,
St. Michael’s Hospital; Institute for Biomedical Engineering Science and Technology (iBEST), Ryerson University
and St. Michael’s Hospital; Department of Biological Sciences, University of Toronto at Scarborough
Rationale and Objectives: The white blood cells defend our body from microorganisms by engulfing the foreign
particles. This defense mechanism is known as phagocytosis. As a result of this defense mechanism, the
hydrogel loaded with therapeutic drugs is usually engulfed by the white blood cells while lowering the efficacy
of the drug delivery methods. Researchers discovered that a worm-like particle with high aspect ratio and a
diameter less than 5 microns is known to significantly improve the delivery of the therapeutic drug.
Methods: With Poly (Ethylene Glycol) Diacrylate (PEGDA 700) and 2- Hydroxy-2-MethylPropiophenone (Darocur
1173), the free radical ions induce the cross-linking of the monomer unit (PEGDA 700) in the presence of UV
light. An array of 15 X 15 circular-shaped photomask was chosen to direct the UV light onto the prepolymer
solution. With a UV exposure time and intensity of 100 ms and 80%, respectively, 215 cylindrical microparticles
were synthesized in one pulse of UV radiation for 100 ms.
Results: With the help of 40X microscope objective (i.e at a higher magnification scale), the worm-like particles
were synthesized with particle diameter and length of 6 μm and 30 μm, respectively. In the presence of acrylic
acid and Rhodamine acrylate in the prepolymer solution, the microparticles were fluorescently labeled and
negatively charged which binds to immunoglobulin antibodies (IgG) to study if the shape of the microparticles
has any effect on the phagocytosis and drug delivery.
Conclusion and Significance: For the first time, we have utilized the free radical polymerization technique to
fabricate the worm-like particles using neutrally charged hydrogel (PEDGA700). This technique allows a high
throughput synthesis of worm-like particles and facilitates mass production of up to 1 million particles
depending on the number of UV pulses per hour.
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2021
Poster #14: Thin ECM-based membrane integration for organ-on-

a-chip applications
Jeremy Newton, Edmond W. K. Young
Department of Mechanical and Industrial Engineering, University of Toronto; Institute of Biomedical

Rationale and Objectives: Thin membranes are important structures in organ-on-a-chip (OOC) devices that
enable precise control of the 3D arrangement of cells. In most devices, synthetic membranes with a biological
coating are used, but recent trends have shown efforts to adapt thick biological gel manufacturing methods to
produce thin, purely biological membranes in OOC devices. Our project aims to develop a simple and user-
friendly method for fabricating a thin collagen-based membrane in a thermoplastic device for air-liquid
interface coculture experiments.
Methods: A micro-milled thermoplastic device was designed to enable patterning of a collagen hydrogel using
a micropipette, which was then air-dried to produce a thin collagen membrane in its place. Confocal reflective
imaging was used to characterize the thickness and fibrous architecture of the membrane. The device was
designed to allow user-friendly cell culture on either side of the membrane without further manipulation after
membrane fabrication. The ability of the device to mimic the thin walls of lung alveoli was tested by culturing
monolayers of epithelial and endothelial cells on opposing sides of the membrane. The integrity of the
monolayers was tested by verifying the presence of key cell junction proteins.
Results: Device design optimization was conducted to improve control of membrane characteristics and enable
robust adhesion of the membrane to the device. An optimized device operation protocol was developed to
avoid trapped air bubbles in the complex geometry of the membrane support structure and to enable
successful co-culture of a human epithelial cell line and human umbilical vein endothelial cells.
Conclusion and Significance: The preliminary characterization of a user-friendly coculture platform utilizing a
purely biological thin cell culture substrate for barrier tissue modelling has been completed. Future work will
include a demonstration of the physiological relevance of the platform in modelling the alveolar air-blood
barrier, with a focus on medium to high-throughput analysis methods.
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Poster #15: Modeling HFpEF myocardial dysfunction in a heart-

on-a-chip platform
Omar Mourad, Sara S. Nunes
Toronto General Hospital Research Institute; Institute of Biomedical Engineering, University of Toronto;
Laboratory of Medicine and Pathobiology, University of Toronto; Heart & Stroke/Richard Lewar Centre of
Excellence, University of Toronto
Rationale and Objectives: Heart failure with preserved ejection fraction (HFpEF) is an advanced stage of heart
disease characterized by impaired muscle relaxation, a thickened myocardium, and diffuse fibrosis. It is a
systemic disease and is highly associated with comorbidities such as metabolic syndrome. Due to its complex
nature, no in vitro HFpEF models are available yet, and animal models are unsatisfactory. My research is aimed
at generating an in vitro model of HFpEF myocardial dysfunction by leveraging established heart-on-a-chip
technology used by our group.
Methods: In our heart-on-a-chip platform, human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are
combined with cardiac fibroblasts in a fibrin gel to generate microtissues that self-assemble around flexible
rods. Rod deflection can be measured to determine passive and active force of these beating tissues. To model
the HFpEF myocardium, we will design a treatment protocol that results in tissues that display increased
passive force with an unchanged active force relative to healthy tissues. To achieve this, we will treat the tissues
with combinations of factors known to induce specific phenotypes associated with HFpEF such as hypertrophy
and fibrosis, as well as an environment that mimics metabolic syndrome.
Results: I have screened combinations and doses of factors known to induce hypertrophy and fibrosis of CMs in
vitro in monolayer and identified a combination that robustly increases hPSC-CM cell size. In addition, qPCR
experiments have confirmed that the hypertrophy that I observed is pathological and not physiological,
evidenced by upregulation of fetal genes. I have used this treatment regimen in on our heart-on-a-chip
platform and generated tissues with 2-3x increased passive force, and unchanged active force relative to
control.
Conclusion and Significance: These studies will enable us to uncover molecular pathways that might be
involved in the pathophysiology of HFpEF myocardial dysfunction. An in vitro model of the disease can be used
as a drug screening platform in the future, which is highly significant due to the lack of empirical therapeutic
options for HFpEF currently available.
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2021
Poster #16: Microfluidic device to quantify SARS-CoV-2

neutralizing antibodies
Md. Almostasim Mahmud, Liangcheng Henry Xu, Dustin J. Little, Darius G. Rackus, Scott Tsai
Department of Chemistry and Biology, Ryerson University; Institute for Biomedical Engineering, Science and
Technology; Department of Mechanical and Industrial Engineering, Ryerson University
Rationale and Objectives: Generation of herd immunity to SARS-CoV-2 either through vaccination or natural
infection is key to ending the COVID-19 pandemic. It is important to establish strong and effective serological
surveillance through the measurement of immunity in a population to direct public health policy including plans
for consecutive vaccine doses.
Methods: Individual immunity to SARS-CoV-2 is defined by the presence of virus specific neutralizing antibodies
(nAbs). These antibodies prevent the virus from entering host cells by occluding access to a specific host
receptor angiotensin-converting enzyme 2 (ACE2). Based on the same principle, we are developing a paper-
based microfluidic device that can test the level of nAbs present in a blood sample. Upon loading, the sample
is mixed with pre-spotted recombinant SARS-CoV-2 Spike glycoprotein receptor binding domain (RBD)
conjugated to a gold nanoparticle (GNP) label.
Results: Several test dots of ACE2 are spotted along the detection zone with increasing concentrations to
implement a semi-quantitative gradient assay. Counting the number of developed spots is inversely
proportional to the concentration of nAbs present in the sample. So far, we are able to demonstrate semi-
quantitative measurements of neutralization using soluble ACE2 or nAbs as the neutralizing agent.
Conclusion and Significance: At this stage, the device requires further optimization to increase the sensitivity of
the device, to get consistent test readout for standard samples, and to increase shelf life. After optimization,
this rapid test device can become a convenient and user-friendly platform for SARS-CoV-2 serology testing.
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2021
Poster #17: Single-cell approaches to studying bacterial

competition
Tianyi Ma, Daniel Nino, Josh Milstein
Department of Chemical and Physical Sciences, University of Toronto; Department of Physics, University of
Toronto
Rationale and Objectives: The fate of an isogenic bacterial population is related to single-cell phenotypes and
the individual behavior can lead to complex population dynamics. However, the phenotypic heterogeneity of
bacteria is complex since it responds to a manifold of biological and environmental factors. It is challenging to
study cell-to-cell variations in behavior that are involved in the bacterial competition since conventional bulk
methods lack sufficient control of the environmental conditions and do not provide single-cell information.
Methods: Microfluidic monolayer chambers (MMC) are designed to couple with time-lapse fluorescence
microscopy and are able to provide the necessary spatiotemporal resolution for investigating bacterial
competition at a single-cell level. Moreover, a single-cell analysis essay has been developed to extrapolate the
population dynamic and single-cell information during the competition.
Results: We have been studying the mechanisms that enable one of the micro-colonies of two bacterial strains
(E. coli strains pBAD-eGFP and pBAD-mCherry) to dominate within a small, structured space, mimicking the
onset of infection and/or providing a minimal/toy model of the microbiome. Single-cell analysis on various
aspects of bacterial heterogeneity that are related to spatial competition, such as fixation time and cellular
organization, have been studied.
Conclusion and Significance: The competition configurations, as well as the fixation time, are shown to be
dependent on the spatial structure of the confined microfluidic chamber. It emphasizes the necessity of
considering the contribution of spatial conditions during microbial colonization and interaction.
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2021
Poster #18: Early indicators of a “zombie” microfluidic devices

Jurgen Fresheri, Lorenzo Gutierrez
Microfluidic Bioservices Department, StarFish Medical
Rationale and Objectives: Despite their initial appearance microfluidic device prototypes often fail during use,
so called “zombie” prototypes. Early design strategies and techniques can significantly increase the likelihood
of functional success and increase production yield of the device. The objective of this study is to determine
and mitigate factors contributing to the high rate of failure during the prototyping process. Specifically, we
would like to establish testing methods and procedures to reduce cartridge failure and increase yield.
Methods: A representative microfluidic device shall be prototyped and detailed drawings shall outline critical
features such as: channel dimensions, reservoirs, mixing chambers, detection area, and off-the-shelf component
interfaces. These features shall be machined by a CNC micro-mill with channels as small as 50um wide. The
prototyped device shall consist of a stack-up of at least four layers bonded by an automated adhesive
dispenser and cured for 24 hours. A batch of 5 microfluidic devices shall be built and subjected to detailed
optical inspection using a stereo microscope and profilometer. Basic functionality will be assessed using
pressure decay and fluidic flow tests. Specific functionality of the device shall be assessed by running an assay
using a defibrinated sheep blood sample.
Results: Test results showed that all the units in the batch passed optical inspection, pressure decay and fluidic
flow tests. However, only one unit successfully ran the assay. In most cases, the devices failed before
completion of the assay. The study showed the following observations: (1) Missing parts or features which are
critical to the test, (2) The device design was too complex which contributed to the potential failure mode, (3)
Incompatibility of the device material with assay reagents, (4) Inadequate integration of off-the-shelf
components, (5) Tolerance issues.
Conclusion and Significance: The goal in microfluidic prototyping is to produce a device that has reliable
functionality and high production yield. A lack of controls during the fabrication process creates lots of potential
failure modes. The study showed that in every step of the process, the combination of these failure modes can
frustrate troubleshooting and debugging the design and fabrication of the prototype. Mitigating and
decoupling these failure modes can significantly reduce the number of prototype iterations and increase the
reliability and production yield of the device.
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2021
Poster #19: Modelling SARS-CoV-2 infection in vascularized

perfusable system
Rick Xing Ze Lu, Benjamin Fook Lun Lai, Naimeh Rafatian, Dakota Gustafson, Kathryne Howe, Jason Fish, Milica
Radisic
Institute of Biomaterials and Biomedical Engineering, University of Toronto; Toronto General Research Institute;
Department of Chemical Engineering and Applied Chemistry, University of Toronto
Rationale and Objectives: Severe acute respiratory syndrome coronavirus (SARS-CoV-2) is known for its impact
on the respiratory tract. However, recent data suggest that up to 78% of people experiencing COVID-19 have
some cardiac effects, yet, effective medications for the treatment of COVID-19 induced myocarditis is lacking.
In this proposed study, we present a human-specific model composed of cells of human origin to investigate a
novel anti-inflammatory peptide QHREDGS as a potential therapeutic for SARS-CoV-2 induced cardiovascular
complications.
Methods: In our study, we present a human-specific in vitro model composed of cells of human origin. Human
umbilical vein endothelial cells (HUVEC) were cultured in the lumen of the system to represent
microvasculature. The endothelial cells were infected with either NL63 or SARS-CoV-2, and characterized
endothelial barrier function, monocyte recruitment, extensive cytokine and chemokine profile, and the effect of
novel QHREDGS as potential therapeutics.
Results: Our preliminary data suggest the ability of SARS-CoV-2 to cause compromised barrier function as
indicated by an increase of FITC-dextran and disrupted EC junction using VE-cadherin staining. Furthermore,
the level of pro-inflammatory cytokines (GM-CSF, IL-1β, IL-6, MCP-1) and soluble EC activation markers (ET-1,
ICAM-1, Ang-II) was significantly increased. The number of PMBC attachment onto the endothelial cells were
increased compared to the chip without coronavirus infection.
Conclusion and Significance: Together, our data show that an organ-on-a-chip device featuring human cells,
can be used to rapidly identify approved drugs that may be repurposed for pandemic-virus applications in crisis
situations that require the accelerated development of therapeutic interventions.
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Poster #20: Extrusion of perfusable elastomeric tubules

Chuan Liu, Scott Campbell, Milica Radisic
Engineering, University of Toronto; Toronto General Research Institute, University Health Network
Rationale and Objectives: Hollow microfibers have attracted increasing attention due to their promising
perfusability and uses in microphysiological systems. However, many of these microfibers have been made of
biomaterials that lack physiologically relevant elasticity, such as alginate. This hinders their ability to mimic the
properties of vessels or tissues in vivo. Thus, this study aims to fabricate microfibers using a synthetic
bioelastomer, poly(itaconate-co-citrate-co-octanediol) (PICO), in order to develop biomimetic models using
high throughput techniques.
Methods: The elastomeric microfibers are fabricated using a coaxial needle attached to the print head of a 3D
printer to control the movement of the needle in the x and y directions. The inner needle is filled with a
sacrificial material (Pluronic F127) in the form of a gel whereas the outer needle is filled with the PICO
prepolymer. As both phases are extruded from the coaxial needle, a tubular structure is formed and collected
in a pluronic solution before UV curing. Intact lumen is revealed after removing the pluronic inside the fiber.
Results: The dimensions of the microfibers can be tuned by varying various printing parameters, including the
ratio between the inner and outer flow rate, the overall flow rate, UV curing time, and needle speed and height.
The resulting tubes are shown to be capable of perfusing fluorescently-labelled beads and their lumen can be
seeded with endothelial cells. Porogen can be introduced into the polymer phase to allow for the potential for
diffusion across the tubular structure, however we are investigating alternative, high throughput methods to
introduce holes into the printed elastomeric tubes.
Conclusion and Significance: Successful coaxial extrusion of elastomeric microfibers that can more effectively
mimic the properties of native vasculature can afford significant improvements in the development of organ-on-
chip systems, particularly if these microfibers can be produced alongside the rest of the device in a high
throughput manner.
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Poster #21: Melt-electrospinning based 3D printing of

polycaprolactone
Jianfeng Li, Hend Kholeif, Ka My Dang, Michael G. K. Brunk, Wesley D. Sacher，Joyce K. S. Poon
Department of Nanophotonics, Integration, and Neural Technology, Max Planck Institute of Microstructure
Physics; Department of Electrical and Computer Engineering, University of Toronto; Karlsruhe School of Optics
and Photonics, Karlsruhe Institute of Technology
Rationale and Objectives: As a Food and Drug Administration (FDA) approved biocompatible material,
polycaprolactone (PCL) is useful for various biomedical applications. Our current study focuses on controlling
the melt-electrospinning (MES) based 3D printed PCL scaffold morphology with different printing parameters
and apply the customized scaffolds towards cell supporting and microdialysis.
Methods: In our investigation, we analyzed the influence of the parameters: working distance, printing speed,
voltage and pressure, on the printed scaffold morphology, specifically to further improve the control over fiber
thickness, fiber alignment, pore size and surface morphology in order to subsequently optimize our scaffold
designs.
Results: Our thorough investigation of the 3D printing parameters allows precise control over MES printed PCL
scaffold morphology per to the requirement of different applications. With the developed method, the follows
have been achieved: (1) PCL fiber diameter can range from 1 to 70 μm; (2) PCL fibers in the scaffold can be
aligned, twisted and a combination of both; (3) Pore size within the printed scaffold can be as small as 2 μm; (4)
With modified design, a single printed PCL layer can have 3D features, which could be beneficial for cell
supporting; (5) With different morphology, cell behavior can be tuned accordingly.
Conclusion and Significance: To mimic native counterpart, a steady and thorough optimization of 3D printed
scaffolds for brain research and treatment is compulsory and improvement of these related parameters should
also be studied. Here, we investigated the influence of different parameters for the MES based 3D printed PCL
scaffolds to create new biocompatible structures for future research.
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Poster #22: Recapitulating pancreatic tumor microenvironment

with organ-on-chip platform
Benjamin Lai, Rick Lu, Yangshuo Hu, Locke Davenport Huyer, Wenkun Dou, Erika Wang, Nikolina Radulovich,
Ming Tsao, Yu Sun, Milica Radisic
Institute Biomaterials and Biomedical Engineering, University of Toronto, Department of Chemical Engineering
and Applied Chemistry, Toronto General Research Institute; Princess Margaret Cancer Centre, University Health
Network; Department of Mechanical and Industrial Engineering, University of Toronto
Rationale and Objectives: Tumor progression relies heavily on the interaction between the neoplastic epithelial
cells and their surrounding stromal partners. This cell cross-talk affects stromal development, and ultimately the
heterogeneity impacts drug efflux and efficacy.
Methods: To mimic this evolving paradigm, we have micro-engineered a three-dimensional (3D) vascularized
pancreatic adenocarcinoma tissue in a tri-culture system composed of patient derived pancreatic organoids,
primary human fibroblasts and endothelial cells on a perfusable InVADE platform situated in a 96-well plate.
Uniquely, through synergistic engineering we combined the benefits of cellular fidelity of patient tumor derived
organoids with the addressability of a plastic organ-on-a-chip platform.
Results: Validation of this platform included demonstrating the growth of pancreatic tumor organoids by
monitoring the change in metabolic activity of the tissue. Investigation of tumor microenvironmental behavior
highlighted the role of fibroblasts in symbiosis with patient organoid cells, resulting in a six-fold increase of
collagen deposition and a corresponding increase in tissue stiffness in comparison to fibroblast free controls.
The value of a perfusable vascular network was evident in drug screening, as perfusion of gemcitabine into a
stiffened matrix did not show the dose-dependent effects on tumor viability as those under static conditions.
Conclusion and Significance: These findings demonstrate the importance of studying the dynamic synergistic
relationship between patient cells with stromal fibroblasts, in a 3D perfused vascular network, to accurately
understand and recapitulate the tumor microenvironment.
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Poster #23: Oil-free droplet microfluidic generation of hydrogel

spheroids
Jennifer Kieda, Sila Appak-Baskoy, Morteza Jeyhani, Maryam Navi, Katherine Chan, and Scott S. H. Tsai
Graduate Program in Biomedical Engineering, Ryerson University; Department of Mechanical and Industrial
Engineering, Ryerson University; Keenan Research Centre for Biomedical Science, St. Michael’s Hospital;
Institute for Biomedical Engineering, Science and Technology (iBEST)—a partnership between
Ryerson University and St. Michael’s Hospital
Rationale and Objectives: Spheroids are three-dimensional (3D) clusters of cells and are commonly used as in-
vitro tumour models to recapitulate in vivo morphology. A limitation of many existing on-chip platforms for
spheroid formation is the use of cytotoxic organic solvents as the continuous phase in droplet generation
processes. Here, we describe an oil-free droplet microfluidic approach that utilizes an aqueous two-phase
system to generate breast cancer hydrogel spheroids.
Methods: We make stock solutions of dextran containing alginate (DEX–alginate), polyethylene glycol (PEG),
and calcium chloride (CaCl2) dissolved in DMEM growth media. Using pressure pumps, we generate
monodisperse DEX-alginate droplets containing MCF-7 cells in a continuous phase of PEG using a microfluidic
device. Downstream of the device, DEX-alginate droplets are introduced to CaCl2, allowing for crosslinking
between alginate and CaCl2, thereby forming hydrogels. Using tubing connected to the outlet of the device,
we collect generated hydrogels and transfer the hydrogels into a 96-well plate. We use a common
chemotherapy drug, doxorubicin, for all cancer drug testing experiments.
Results: We collect and harvest the hydrogels generated from the flow-focusing microfluidics device for up to
seven days. Over time, cells aggregate within the hydrogels, forming spheroids. We apply five conditions to the
cell-containing hydrogels in drug testing experiments, including an untreated control condition and four drug
concentrations over 48 hours. To test for viability, we use a LIVE/DEAD assay on cells in conjunction with
confocal imaging. With increasing doxorubicin concentration, there is a monotonic decrease in the viability of
cells.
Conclusion and Significance: We describe a droplet microfluidic platform for the generation of hydrogel tumour
spheroids by encapsulating MCF-7 cells in droplets using an all-aqueous system. Our platform offers a
biocompatible alternative to oil-water platforms, making the platform suitable for cancer drug testing and
controlling the concentration of cells in the spheroid and monodispersity.
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Poster #24: Deciphering the genetic signature of pancreatic

endothelial cells
Safwat T. Khan, Sara S. Nunes
Institute of Biomedical Engineering, University of Toronto; Toronto General Hospital Research Institute,
University Health Network; Laboratory Medicine and Pathobiology, University of Toronto; Heart and
Stroke/Richard Lewar Centre of Excellence, University of Toronto
Rationale and Objectives: Decades of studies in the vasculature demonstrate vessels show intricate tissue-
specific structural, physiological and functional heterogeneity. In particular, pancreatic islet ECs (iECs) are
uniquely adapted to allow rapid glucose sensing and homeostatic response by insulin-secreting ß cells. In this
project, we have harnessed the unparalleled transcriptomic resolution provided by single cell RNA sequencing
(scRNAseq) and state-of-the-art RNAseq analysis tools to uncover a unique transcriptomic profile for iECs.
Methods: We have isolated EC sequencing data from published scRNAseq datasets from different human and
murine tissues including pancreas/pancreatic islet and performed transcriptomic meta-analysis using novel
bioinformatics tools to determine an islet-specific EC transcriptomic profile.
Results: In this project, we have uncovered unique gene signatures that define different segments of the
vascular bed in the pancreas. Additionally, we have demonstrated potential lineage relationships between iECs
and other tissue ECs. Finally, we have uncovered unique marker genes for iECs against other tissue ECs, which
are cross-species conserved.
Conclusion and Significance: Further characterization of these genetic signatures will provide novel insights into
iEC specialization in glucose homeostasis. This will help facilitate the development of innovative strategies for
improving islet vascularization post-transplantation to treat diabetes and for generating 3D islet vasculature-on-
a-chip for diabetic disease modelling.
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Poster #25: On-the-fly physical property changes of ATPS on

CMP
Byeong-Ui Moon, Liviu Clime, Javier Alejandro Hernandez-Castro, Daniel Brassard, Christina Nassif, Lidija
Malic, Teodor Veres
Life Sciences Division, National Research Council Canada, Boucherville, Quebec
Rationale and Objectives: Aqueous two-phase systems (ATPS) are formed when two incompatible polymer/salt
solutions are mixed in water, and used in many biological applications. To maximize the range of applications
compatible with ATPS, altering and controlling their physical properties (phase transition, density, osmolality,
viscosity and interfacial tension) are of great importance.
Methods: We demonstrate on-the-fly physical property changes of ATPS in microfluidic devices. The properties
and phases of the ATPS are modulated on-demand using a centrifugal microfluidic device filled with
polyethylene glycol (PEG) and dextran (DEX) solutions. Using the centrifugal force and active pneumatic
controls provided by a centrifugal microfluidic platform (CMP), PEG-DEX mixtures are manipulated and
processed inside simple thermoplastic microfluidic devices.
Results: We show on-chip control over the phase change of ATPS by changing a two-phase solution into a
single phase with the addition of water. Single-phase-to-two-phase transition is also demonstrated by adding
ATPS with higher PEG and DEX concentrations. We also demonstrate a density modulation scheme by
introducing an ATPS solution mixed with sodium diatrizoate hydrate. By adding precisely metered volumes of
water, we spontaneously change the density of the solution on the CMP and show that density marker
microbeads fall into the solution according to their corresponding densities. The measured densities of ATPS
show a good agreement with densities of microbeads and analytical plots.
Conclusion and Significance: We conclude that our achieved results highlight the tremendous potential of
CMPs for performing complex on-chip processing of ATPS. We also anticipate that the proposed ATPS-CMP
may be useful for a wide range of applications in biological sample processing, for example, microparticle-
based plasma protein analysis and blood cell fractionation.
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Poster #26: Two-photon direct laser writing of 3D microscaffolds

for neural engineering
Ka My Dang, Jianfeng Li , Michael G. K. Brunk, Wesley D. Sacher, Joyce K. S. Poon
Department of Nanophotonics, Integration, and Neural Technology, Max Planck Institute of Microstructure
Physics; Department of Electrical and Computer Engineering, University of Toronto
Rationale and Objectives: Neuron cultures assays are useful to study neuronal de- and regeneration. To
manipulate the nervous system, 2D cultures have been commonly used. However, the 2D cultures are
insufficient in studying specific physiological features due to the limitation of cell-extracellular matrix
interaction. Conversely, 3D in-vitro culture of neuronal cells provides a more complex environment to improve
tissue organization. Therefore, 3D culture scaffolds can be a promising approach for organ-on-a-chip platform
and a complement method to animal models.
Methods: We present the fabrication of 3D scaffolds using two-photon direct laser writing for spatial
organization of neuron-like PC12 cells. Two-photon polymerization (2PP) is a technique using laser writing of
biocompatible materials to produce miniaturized 3D scaffolds with nano-and micro-meter features sizes. The
system focuses femtosecond pulses of infrared laser light in the volume of a photosensitive material to write 3D
microstructures. To check the survival of cells cultured on the 3D microstructures, live-dead assays have been
performed on the PC12 neuronal cell cultures. The viability of the cells has been measured by imaging with a
confocal microscope.
Results: To identify dimensions suitable for healthy cell development and the connection of neuronal networks
on 3D scaffolds, hexagonal and rectangular structures with different pore sizes between 20 and 50 µm have
been fabricated and investigated. The results show that PC12 neuronal cells adhere well on the 3D scaffolds
and the cells can connect to form 3D networks.
Conclusion and Significance: We presented an approach to fabricate 3D microstructures precisely using two-
photon direct laser writing of biocompatible materials. Neuronal cell growth has been studied on the 3D
microstructure, revealing healthy proliferation on the scaffolds with a small number of dead cells. The produced
scaffolds can be structured with the purpose of guiding neuronal outgrowth.
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Poster #27: Development of a delivery platform for protein-based

therapies
Shirley Chung, Jason T. Maynes, J. Paul Santerre
Rogers Centre for Heart Research, University of Toronto; Program in Molecular Medicine, SickKids Research
Institute; Department of Anesthesia and Pain Medicine, Hospital for Sick Children; Department of
Anesthesiology and Pain Medicine, University of Toronto; Faculty of Dentistry, University of Toronto
Rationale and Objectives: Leading-edge regenerative therapies targeting heart failure have utilized the delivery
of cardiac progenitor cells (CPC) to improve heart function. However, recent work has shown that the cells
themselves may not be the main therapeutic vector, but rather the components that the cells secrete (the
secretome) provides significant beneficial effects. We hypothesize that D-PHI based nanoparticles (NPs) will
deliver therapeutic components of CPC secretomes through a release profile that will induce reparative
function in damaged heart tissue.
Methods: The research team has synthesized a degradable polar hydrophobic ionic polyurethane, referred to
as D-PHI, that has previously been used to generate cardiac cell-compatible nanofibers and NPs that can be
employed as a delivery vehicle. Both modalities will be explored to achieve effective delivery of secretomes
components to the myocardium. D-PHI NPs were fabricated using an emulsion inversion polymerization
technique and characterized through dynamic light scattering and NanoSight to determine the diameter and
zeta potential before and after coating with our proteins. A WST-1 assay using human cardiac fibroblasts was
performed to determine the cell compatibility of our NPs.
Results: Early work shows that D-PHI NPs have an average diameter of 207 ± 45 nm (PDI < 0.2) and a zeta
potential of -70 ± 3.4 mV. WST-1 results show D-PHI NPs were biocompatible with up to 2 x10^10 NPs/mL.
Additionally, the Maynes Lab has defined a set of proteins that are enriched in CPC secretomes. These proteins
will be loaded onto D-PHI NPs via charge-driven surface coating. Currently, two proteins identified to be of
interest have been successfully loaded onto the NPs. Changes in the zeta potential and diameter of the NPs,
when compared to the uncoated NPs, are used as early indicators of successful protein binding (protein 1
coated NPs: 234 ± 55 nm, -39 ± 6.5 mV; protein 2 coated NPs: 230 ± 47 nm, -52 ± 2.6 mV).
Conclusion and Significance: In conclusion, biocompatible D-PHI NPs were fabricated using an inversion
emulsion polymerization technique and therapeutically relevant proteins identified by the Maynes Lab were
subsequently coated onto the NPs. The delivery of these proteins to damaged myocardium has the potential to
induce reparative function in damaged heart tissues.
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Poster #28: Microfluidic platform for intracellular phase

separation experiments
Katherine Chan, Maryam Navi, Jennifer Kieda, Scott S. H. Tsai
Graduate program in Biomedical Engineering, Ryerson University; Department of Mechanical and Industrial
Engineering; Keenan Research Center for Biomedical Science, St. Michael’s Hospital; Institute for Biomedical
Engineering, Science, and Technology (iBEST)
Rationale and Objectives: Cells organize their contents into droplet compartments, termed coacervates, in a
process called liquid-liquid phase separation (LLPS). Characterizing coacervate composition and formation is
necessary to understand their suggested functions in mediating viral-host interactions and neurodegenerative
disease. However, current methods to understand modulators of coacervation require extensive manual sample
preparation. We present a microfluidic platform that eliminates manual preparation for high-throughput,
minimal reagent in vitro experiments.
Methods: We eliminate the extensive micro-pipetting that is characteristic of conventional methods by

preparing samples in a double-layer, polydimethylsiloxane-on-glass microfluidic chip. In the bottom layer, a
diffusive gradient mixer produces numerous solutions of varying concentrations. Then, a phase separation
trigger in the top layer is introduced to the bottom layer via connected inlets. The dual solutions are mixed and
observed for phase separation in reservoirs, where each reservoir contains samples with specific compositions.
By preparing samples simultaneously, this microfluidic platform facilitates the collection of several data points in
a single experiment, with the potential for further scale up.
Results: We construct a phase diagram of an aqueous two-phase system (ATPS) of polyethylene glycol (PEG)
and dipotassium phosphate (K2HPO4). The bottom layer creates PEG samples of varying weight percent. Upon
introduction of K2HPO4 from the top layer to the bottom layer, the solutions mix or phase separate. Then, we
construct a phase diagram describing coacervation of Mycobacterium tuberculosis phase-separating protein
Rv1747 and PEG, the phase separation trigger. An array of varying concentrations of Rv1747 is generated in the
bottom layer of the device. Following the introduction of PEG from the top layer, samples form a
homogeneous solution or phase-separated coacervates.
Conclusion and Significance: We present a microfluidic platform that collects data in a shorter time and with
less reagents than conventional methods. We expect this platform to provide information about the conditions
under which phase separation occur so that further conclusions about the functional relevance of phase
separation to pathology can be made.
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Poster #29: Epithelial to mesenchymal transition of epicardial

cells and EMT-on-a-chip
Dawn Bannerman, Simon Pascual Gil, Karl Wagner, Qinghua Wu, Milica Radisic
Chemical Engineering and Applied Chemistry, University of Toronto; Institute of Biomedical Engineering, University of
Toronto; Toronto General Hospital Research Institute, University Health Network
Rationale and Objective: A promising way to repair damaged tissue following a heart attack involves the use of cardiac
patches. Patches are placed on the epicardial surface, which is comprised of a barrier layer of epicardial cells (ECs). ECs
can transform into more motile cell types through a process called epithelial-to-mesenchymal transition (EMT), which if
sufficiently controlled, could improve patch integration. This work examines the effect of bioactive molecules such as
extracellular vesicles (EVs) on EC layers.
Methods: EVs are isolated from human ECs undergoing EMT. These EVs are characterized and applied as a treatment to
cultured EC sheets. EVs released from a controlled delivery matrix are compared with soluble EVs. Transition of the EC
layers into various cells fates is assessed by immunofluorescence staining for the presence of epicardial markers WT1 and
ZO-1, fibroblast marker Vimentin, and smooth muscle marker α-smooth muscle actin. In addition, a custom-designed
“EMT-on-a-chip” device fabricated from polystyrene moulds assembled with cell culture plates provides a tool for
assessment of cell migration and phenotypic changes in response to factors introduced in a directionally dependent
manner.
Results: EVs derived from ECs have been isolated, characterized, and sustained release from a fibrin gel has been
demonstrated over a two-week period. The effect of EV treatments on ECs is under investigation. The EMT-on-a-chip
device has been successfully fabricated. ECs have been cultured in specified regions while biomaterials containing
bioactive molecules are maintained in a separate region with a section bridging the two areas for assessment. The use of
this device for cell migration assays and assessment of phenotypic changes will be completed upon selection of
favourable treatment conditions from the EC treatment experiment.
Conclusions and Significance: The focus of this work is to achieve control the EMT process of ECs in vitro to
support cardiac patch technology. Uncovering treatment conditions that result in favourable EMT pathways of ECs can be
applied to cardiac patch systems to improve patch integration, as well as potentially support reparative processes.
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2021
Poster #30: A microfluidic device for implementing controlled

soft particle interaction
Aysan Razzaghi, Arun Ramachandran
Department of Chemical Engineering and Applied Chemistry, University of Toronto
Rationale and Objectives: In the past, microfluidics has been integrated with external force fields such as
magnetic, electric, acoustic, optical fields to perform separation, adhesion, and manipulation of biological cells
and soft particles. In several cases, however, the use of these external fields is precluded due to the limitations
on the type of material that can be used as the channel or the sample. In this work, we demonstrate the use of
a field naturally available in the microfluidic platform – the hydrodynamic field – to implement the same studies.
Methods: The device employs a microfluidic channel with six inlet/ outlet ports to produce two stagnation
points, which can be used to manipulate two particles simultaneously. We implement an analytical solution of
the flow field in an active feedback loop to continuously update the required flow rates from the ports and steer
the particles independently along arbitrary trajectories. The effects of flow perturbation due to the presence of
the other drop are also accounted for in the analytical solution.
Results: To demonstrate the functionality of the device, we hydrodynamically manipulated two pancake-shaped
perfluorodecalin drops in silicone oil to approach and coalesce with each other either in a head-on or a
glancing configuration at different, adjustable extensional rates. The experimental results of the drainage time
for coalescence show good agreement with a scaling analysis developed for pancake-shaped drops in an
emulsion. The drainage time was found to scale with the capillary number (the ratio of viscous to interfacial
forces), with a pre-factor that depends on the disjoining stresses, extensional rate, viscosity ratio, and the
confinement of the drops.
Significance: While the device has been tested on emulsion drops, by suitably modifying the control algorithm
and the microfluidic fabrication techniques, this device can very well be used to study the interactions between
vesicles, microgels and biological cells.
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2021
Poster #31: Engineering a gingival graft under perfusion

Brian Webb, Paul Santerre
Institute of Biomaterials and Biomedical Engineering, University of Toronto; Faculty of Dentistry, University of
Toronto; Institute of Biomedical Engineering, University of Toronto; Translational Biology and Engineering
Program, Ted Rogers Centre for Heart Research
Rationale and Objectives: There is currently no suitable tissue-engineered-solution for gingival recession, a soft
tissue defect which affects more than half of the US population. The central objective of this work is to show
that seeding and layering electrospun degradable/polar/hydrophobic/ionic polyurethane and polycarbonate-
urethane (D-PHI/PCNU) nanofibrous scaffolds, combined with human adipose-sourced primary microvascular
endothelial cells (HAMVECs) and fibroblast-like human adipose-derived stem cells (ASCs), will generate host-
centric tissue constructs.
Methods: D-PHI/PCNU scaffolds were fabricated via electrospinning membranes and then layered after they
had been seeded with a co-culture of ASCs and human umbilical vein endothelial cells (HUVECs). Future work
will use HAMVECs to replace HUVECs. Preliminary experiments assessed the workflow for layering the
scaffolds, and a robust layering method was developed, where a scaffold layering device was conceived by
laser-cutting a support from polytetrafluoroethylene and was subsequently validated in a second layering
experiment. Hematoxylin and eosin (H&E) staining were used to assess extracellular matrix and cell density.
Scanning electron microscopy (SEM) was used to image the constructs.
Results: ASCs were isolated from host human adipose tissue and the scaffold layering device was validated.
SEM imaging and H&E staining indicate that the co-culture is producing high levels of tissue. Further, Movat
pentachrome staining and immunostaining of the sectioned constructs is ongoing to characterize cell types.
The multilayered constructs will be perfused to catalyze microvasculature formation. A bioreactor has been
designed and fabricated from polycarbonate and will be validated in the near future.
Conclusion and Significance: Tissue-engineered constructs may provide a faster procedure, and less painful
alternative, while lowering the chance of infection when compared to the current treatment of autografts. The
preliminary results look promising for the development of an autologous gingival graft, with encouraging
potential for translation into clinical practice.
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Participating Companies
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VIRTUAL

Winners of Microfluidics Art Competition

First place: Rick Lu, PhD Candidate, Institute of Biomedical Engineering,

Image description: Confocal microscopy image of a vascularized tissue in

Second place: Siwan Park, PhD Candidate, Institute of Biomedical

Image description: Pseudo-coloured SEM image of a carbon black

Third place: Zongjie Wang, PhD Candidate, Edward S. Rogers Sr.

Image description: SEM image revealing the 3D-printed

Faculty of Applied Science & Engineering, University of Toronto

Faculty of Arts & Science, University of Toronto

Leslie Dan Faculty of Pharmacy, University of Toronto

Temerty Faculty of Medicine, University of Toronto

National Science and Engineering Research Council of Canada

Dear symposium attendees and distinguished guests,

Axel Guenther and Teodor Veres

9:00-9:45 Biofabrication keynote (live)

9:45-10:00 – MOVEU MOVEMENT BREAK –

10:00-10:45 Selected biofabrication presentations* (10-min recorded presentations followed by

11:00-12:00 Poster session 1

12:00-1:00 – LUNCH BREAK –

1:00-2:30 CRAFT session: updates, new facilities and projects

2:30-2:45 – MOVEU MOVEMENT BREAK –

3:30-4:30 Industry exhibit hall

August 26: TOeP and Organ-on-a-Chip

9:45-11:15 Trainee networking event

11:30-12:15 Selected TOeP trainee presentations (10-min recorded presentations followed by 5-

12:15-1:15 - LUNCH BREAK -

1:15-2:15 Poster session 2

2:15-3:00 Selected organ-on-a-chip presentations* (10-min recorded presentations followed by

August 27: Diagnostics

9:45-10:45 Selected diagnostics presentations* (10-min recorded presentations followed by 5-

11:00-12:00 Industry exhibit hall

12:00-1:00 - LUNCH BREAK –

Engineering epithelial organoids-on-a-chip

Scheduled for: August 25, 2021 from 9:00-9:45

Jason A. Burdick is the Robert D. Bent Professor of Bioengineering at the University of

Microfluidics-generated microgels for the fabrication of biomedical

Scheduled for: August 26, 2021 from 9:00-9:45

Yoon-Kyoung Cho is currently a full professor in Biomedical Engineering at UNIST and a

Lab-on-a-disc for the detection of rare cells and extracellular

Scheduled for: August 27, 2021 from 9:00-9:45

The Pandemic Preparedness Program: Analytical validation and

Scheduled for: August 27, 2021 from 2:00-2:45

Actuation of three-dimensional-printed nanocolloidal hydrogel

Department of Chemistry, University of Toronto

Methods: Hydrogel sheets were composed of a suspension of cellulose nanocrystals(CNC),

Scheduled for: August 25, 2021 from 10:00-10:15

*Competing in CRAFT People’s Choice Award

Effects of sodium ascorbate on the deposition of extracellular

Institute of Biomedical Engineering, University of Toronto; Department of Pharmacology and Toxicology,

Scheduled for: August 25, 2021 from 10:15-10:30

*Competing in CRAFT People’s Choice Award

One-step formation of protein-based tubular structures for

Methods: The extrusion-based bioprinting approach is enabled by a microfabricated multi-layer PDMS

Scheduled for: August 25, 2021 from 10:30-10:45

*Competing in CRAFT People’s Choice Award

Investigating the role and mechanisms of extracellular vesicle

Scheduled for: August 26, 2021 from 11:30-11:45

Development and characterization of vessel arteriovenous

Scheduled for: August 26, 2021 from 11:45-12:00

Microfluidic arrays of dermal spheroids: a throughput screening

Department of Chemistry, University of Toronto; Department of Mechanical and Industrial Engineering,