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3234/6394192 by Istituto di Matematica Applicata e Tecnologie Informatiche "Enrico Magenes" del CNR user on 28 March 2023
3234 Basic Science – Cardiac Biology and Physiology, Biomaterials, Tissue Engineering

Enhancement of engineered cardiac tissues by promotion of hiPSC-cardiomyocyte proliferation

R. Hesselbarth 1 , T.U. Esser 1 , K. Roshanbinfar 1 , S. Struefer 2 , D.W. Schubert 2 , F.B. Engel 1
1
University hospital Erlangen, Erlangen, Germany; 2 Friedrich Alexander University, Erlangen, Germany
Funding Acknowledgement: Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Deutsche
Forschungsgemeinschaft

Background/Introduction: Cardiac tissue engineering is a promising
strategy to generate human cardiac tissues for modelling cardiac diseases,
screening for therapeutic drugs, and repairing the injured heart. Yet, sev-
eral issues remain to be resolved including the generation of tissues with
high cardiomyocyte density.
Purpose: Determining the effects of the induction of human-induced
pluripotent stem cell-derived (hiPSC) cardiomyocyte proliferation post-
fabrication.
Methods: hiPSCs were differentiated into cardiomyocytes, embedded with
or without CHIR990121 at three concentrations in a collagen pre-gel, and
cast. The engineered cardiac tissues were then cultured in the absence
or presence of CHIR99021 for up to 35 days. Hydrogels and engineered
cardiac tissues were analysed utilizing rheology and assays to determine
viability, proliferation, calcium flow, and contractility.
Results: Here, we show that the integration of CHIR99021 in collagen I
hydrogels promotes proliferation of hiPSC-cardiomyocytes post-fabrication
improving contractility of and calcium flow in engineered cardiac tis-
sues. Presence of CHIR99021 has no effect on the gelation kinetic or
the mechanical properties of collagen I hydrogels. Analysis of cell den-
sity and proliferation based on Ki-67 staining indicates that integration
of CHIR99021 together with external CHIR99021 stimulation increases
hiPSC-cardiomyocyte number by ∼2-fold within 7 days post-fabrication.
Analysis of the contractility of engineered cardiac tissues after another
3 days in the absence of external CHIR99021 shows that CHIR99021-
induced hiPSC-cardiomyocyte proliferation results in synchronized calcium
flow, rhythmic beating, increases speed of contraction and contraction am-
plitude, and reduces peak-to-peak time. The CHIR99021-stimulated engi-
neered cardiac tissues exhibited spontaneous rhythmic contractions for at
least 35 days.
Conclusion: Collectively, our data demonstrate the potential of induced
cardiomyocyte proliferation to enhance engineered cardiac tissues by in-
creasing cardiomyocyte density and reducing arrhythmia.

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