Definition of cell culture

Cell culture refers to the removal of cells from

an animal or plant and their subsequent
growth in a favorable artificial environment.
 Terminology
• Organ culture vs. cell culture
• Primary Culture (heterogeneous population of cells)
• Confluency
• Sub culture
• Passage number
• Clone
• Cell strain
• Cell Line (Finite vs. Continuous Cell Line)
Trypsinization
 Growth curve
• Doubling time
• Population doubling
 Terminology
• Organ culture vs. cell culture
• Primary Culture (heterogeneous population of cells)
• Confluency
• Sub culture
• Passage number
• Clone
• Cell strain
• Cell Line (Finite vs. Continuous Cell Line)
 Types of cells
• On the basis of morphology
– Fibroblast like- cells attached to an substrate Adherent cells in
– Epithelial like-attached to a substrate monolayer culture

– Lymphoblast like- cells do not attach Suspension culture

• On the basis of differentiation state

– Differentiated (chondrocytes, osteoblasts, hepatocyte,…)
– Undifferentited (stem cells)
Epithelial like
 Types of cells
• On the basis of morphology
– Fibroblast like- cells attached to an substrate
– Epithelial like-attached to a substrate
– Lymphoblast like- cells do not attach
• On the basis of differentiation state
– Differentiated (chondrocytes, osteoblasts, hepatocyte,…)
– Undifferentited (stem cells)
 What is a stem cell?
stem cell

SELF-RENEWAL DIFFERENTIATION
(copying) (specializing)

specialized cell
stem cell
e.g. muscle cell, nerve cell
 Why self-renew AND differentiate?
1 stem cell

1 stem cell 4 specialized cells

Self renewal - maintains Differentiation - replaces dead or damaged
the stem cell pool cells throughout your life
 Self Renewal (Regeneration)
• Stem cells are
capable of
dividing &
renewing
themselves for
long periods
 Two types of stem cells
• Embryonic Stem Cells (ESC): received from:
– Embryos created in vitro fertilization
– Aborted embryos
• Adult Stem Cells (ASC): can be received from:
– Limited tissues (bone marrow, adipose tissue, muscle,
dental pulp, amnion, synovial membrane)
– Placental cord
– Dental pulp
Embryonic and adult stem Cell
 Embryonic vs Adult Stem Cells
• Totipotent • Multi or pluripotent
– Differentiation into ANY – Differentiation into some
cell type cell types, limited outcomes
• Large numbers can be • Limited numbers, more
harvested from embryos difficult to isolate
(Legal issue ) • Less likely to cause
• May cause immune immune rejection, since
rejection the patient’s own cells can
– Rejection of ES cells by be used
recipient has not been
shown yet
• Teratoma
Understanding tumorigenic potential
of pluripotent stem cells

(Cell Cycle. 2009;8(16): 2608-2612).

 Hematopoietic stem cells

Human macrophages were derived from

hematopoietic stem cells in the bone marrow.
Mesenchymal stem cells

J Anat. Feb 2004; 204(2): 133–139.

Neural stem cell
 Potential Uses of Stem Cells
• Basic research
– Human development
– molecular basis of cancer
– Molecular mechanisms for gene control
– Role of signals in gene expression &
differentiation of the stem cell
– Signaling pathways
• Biotechnology
– drug discovery
• Cell therapy
Potential Uses of Stem Cells
 Cell biology methods
• Transfection
– GFP
– siRNA
• Live cell imaging
•Chemotaxis
•Angiogenesis
•Migration / Wound healing
•Cell Culture Under Flow
• Real time RT PCR
• Cytological methods ( H&E, …)
Transfection
Recombinant DNA
 Cell biology methods
• Transfection
– GFP
– siRNA
• Live cell imaging
•Chemotaxis
•Angiogenesis
•Migration / Wound healing
•Cell Culture Under Flow
• Real time RT PCR
• Cytological methods ( H&E, …)
Mechanical shear stress

Red – F-Actin (Phalloidin-Alexa 633)

Green – VE-Cadherin (VE-Cadherin (D87F2) XP)
Blue – Cell nuclei (DAPI)
 Articular cartilage

Healthy Joint Trumatic injury

Osteoarthritis

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 Cartilage tissue engineering

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 Scaffolds

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