1. Types of Cell Culture

2. CELL CULTURES, CELL LINES & CELL STRAIN
Isolated Tissue
Cell or tissue culture in vitro
Primary culture
Sub-culture
Secondary culture
Sub-culture
Cell Line
Single cell isolation
Successive sub-culture
Immortalization
Loss of control
of cell growth
Senescence
Cloned cell line
Transformed cell line
Cell Strain

3. CULTURING CELLS IN LABORATORY
Revive frozen cell population
Isolated from tissue
Maintain in culture (aseptic cinditions)
Typical
cell culture flask
Sub-culture (passaging)
‘Mr Frosty’
Used to freeze cells
Cryopreservation

4. DOUBLING TIME : Time taken by cells to double the population.
GENERATION TIME : The number of times cell population has doubled itself.
DOUBLING TIME : Time taken by cells to double the population.
CELL LINE : When the primary culture is first sub-cultured or passaged.
CELL STRAIN : When a particular type of cell lineage is selected, characterized and cloned it is called a cell strain.
PASSAGE NUMBER : The number of times a culture has been sub-cultured.

5. Types of cell cultured Primary cultures
Derived directly from animal tissue,
embryo or adult.
Cultured either as tissue explants or single cells.
Initially heterogeneous – containing various types of cells.
Finite life span in vitro.
Retain differentiated phenotype.
Mainly anchorage dependant.
Exhibit contact inhibition.

6. Types of cell cultured Secondary cultures
Derived from a primary cell culture.
Isolated by selection or cloning.
Becoming a more homogeneous cell population that is contains a specific cell type.
Finite life span in vitro.
Retain differentiated phenotype.
Mainly anchorage dependant.
Exhibit contact inhibition.

7. Types of cell cultured Continuous cultures
Derived from a primary or secondary culture
Immortalised:
Spontaneously
By transformation
Serially propagated in culture showing an increased growth rate
Homogeneous cell population
Loss of anchorage dependency and contact inhibition
Infinite life span in vitro
Differentiated phenotype:
Retained to some degree in cancer derived cell lines
Very little retained with transformed cell lines
Genetically unstable

8. Check confluency of cells Transfer to culture flask
PASSAGING
NEED TO PASSAGE CELLS:-
To maintain cells in culture.
To increase cell number for experiments/storage
Check confluency of cells
Remove spent medium
Checking confluency of cells
Wash with PBS
70-80% confluence
Incubate with
trypsin/EDTA
Resuspend in serum
containing media
100% confluence
Transfer to culture flask