2. Types of culture techniques
• 1. Organ culture: Blood allow to clots in a
cavity and cells are grown on them.
• 2. Explant culture.
• 3. Disaggregation of tissue and get individual
cells
3. Cell cultures are described based on two
parameters
• Origin of the cells
– Primary cell culture (from animal or plant tissue)
– Extended culture (multipassage culture – cell
strain)
– Established (transformed) cell lines
• Manner of growth
– Suspension
– Adherent
4. Primary cell cultures
• Cells derived directly from a tissue
• Limited growth potential
• Limited life span
– Usually cell dies after 50 divisions
• Cells are used within days from culture
• May give rise to a cell strain or be
immortalized
• Strain - a lineage of cells originating from
one primary culture
5. Transformed cells, cell lines
• Derived from tumors
• Arise spontaneously in culture
• Primary cells transformed with viruses,
radiation etc
• Grow indefinitely in culture
• Usually unstable complement of chromosomes
• Cell line - cells derived from a single parental
transformed cell
6. Transformed cell lines
• These types of cells do not age in culture
• They are ‘immortal’
• They often lose contact inhibition
• They often lose many normal
characteristics
• They are not dependent on growth factors
• They may express ‘large T-antigen’ a p53
inhibitor
7. Transformation
Characteristics
– Infinite life span
– High growth
potential
– Low growth factor
dependence
– Suspension growth
– Aneuploid
Methods
– Mutagens
– Viruses
– Oncogenes
– Spontaneous
tumors
Not all transformed cell lines can form
tumors, but all tumors contain transformed
cells
11. Classification of cell cultures based on
manner of growth
• Suspension cells
– Grow floating in culture medium
– Cells survive and proliferate without the attachment to culture
container
– Cells cultured from blood, spleen and bone marrow
• Adherent cells
– Grow as a monolayer attached to the surface of culture container
– Cells derived from ectodermal and endodermal embryonic layers
– Fibroblasts, epithelial cells, neurons
– Most of the time require the specialized surface
12. Name Species and tissue of origin Morphology
MRC-5 Human lung Fibroblast
HeLa Human cervix Epithelial
VERO
African Green Monkey
Kidney
Epithelial
NIH 3T3 Mouse embryo Fibroblast
L929 Mouse connective tissue Fibroblast
CHO Chinese Hamster Ovary Fibroblast
BHK-21 Syrian Hamster Kidney Fibroblast
HEK 293 Human Kidney Epithelial
HEPG2 Human Liver Epithelial
BAE-1 Bovine aorta Endothelial
Adherent cell lines
13. Name
Species and tissue of
origin
Morphology
NSO Mouse myeloma Lymphoblastoid-
like
U937
Human Hystiocytic
Lymphoma
Lymphoblastoid
Namalwa Human Lymphoma Lymphoblastoid
HL60 Human Leukaemia
Lymphoblastoid-
like
WEHI 231 Mouse B-cell Lymphoma Lymphoblastoid
YAC 1 Mouse Lymphoma Lymphoblastoid
U 266B1 Human Myeloma Lymphoblastoid
SH-SY5Y Human neuroblastoma Neuroblast
Suspension cell lines
15. Epithelial cells: Spherical or oval, adherent, confluent monolayer is farmed
After sub culturing. Strong cell adhesion and needs extracellular matrix
Like dextrin, collagen, fibrin to serve as support for cell to attach and multiply.
Mouse and rabbit epethilial cell lines are available from cell culture centres.
Endothelial cells: Oval shaped with elongated connective fibres appear like
spines
Embedded in fibronectin network. Farm partial confluent monolayer after
subculturing.
Fibroblast cells: Rich in actin and mayosin febrile proteins. Cells appear slender
and spiny with large vacuolar space between the fibrin network. Adherent cell
type.
Tissue specific neuronal, hepatic, pancreatoc acinar cell lines also available.
Cells from spleen (lymphocytes), blood (RBC, WBC,Platelets) have their own
morphology specific to their metabolic requirements.
16. • In some instances cell cultures may
grow as semi-adherent cells e.g. B95-8
where there appears to be a mixed
population of attached and suspension
cells.
• For these cell lines it is essential that
both cell types are subcultured to
maintain the heterogeneous nature of
the culture.