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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)
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)
• 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)
Types of cells
Adherent cells in monolayer culture
Suspension culture
• 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)
Types of cells
stem cell
What is a stem cell?
stem cell
SELF-RENEWAL(copying)
specialized celle.g. muscle cell, nerve cell
DIFFERENTIATION(specializing)
1 stem cell
Self renewal - maintains the stem cell pool
4 specialized cells
Differentiation - replaces dead or damagedcells throughout your life
Why self-renew AND differentiate?1 stem cell
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 vs Adult Stem Cells
• Totipotent– Differentiation into ANY
cell type
• Large numbers can be harvested from embryos (Legal issue )
• May cause immune rejection– Rejection of ES cells by
recipient has not been shown yet
• Teratoma
• Multi or pluripotent– Differentiation into some
cell types, limited outcomes
• Limited numbers, more difficult to isolate
• Less likely to cause immune rejection, since the patient’s own cells can be used
Human macrophages were derived from hematopoietic stem cells in the bone marrow.
Hematopoietic stem cells
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
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, …)
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)
31
Articular cartilage
Healthy Joint Healthy Joint Trumatic injury Trumatic injury
OsteoarthritisOsteoarthritis