PREREQUISITES

FOR ANIMAL CELL

CULTURE

Asad Naz
Minimum requirements
for growing cell
 Media
 Support survival and growth

 Natural Media
 Artificial Media
Minimum requirements
for growing cells
 Choice of Media
 Type of cell culture (Normal, Immortalized)
 Object of culture (Growth, survival,
differentiation)
Natural Media

 Clots
 Plasma clots (male fowl)

 Biological Fluids
 Amniotic, Ascitic fluid, serum etc

 Tissue extracts
 Chicken and bovine embryo extracts
Artificial Media
 Immediate Survival
 Prolonged Survival
 Indefinite growth
 Specialized function
Classes
 Serum containing media
 Serum free media
 Chemically defined Media
 Protein free media
Serum Containing
Media
 EMEM with 5-20% serum
 Provide plasma protein, peptides, lipids
carbohydrates, minerals and enzyme
 Hormones (cortisone, insulin and testosterone and
prostaglandin)
 Growth factors (PDGF, TGF-p, epidermal growth factors etc)
 Supply protein (fibronectin, spreading factor
Serum Containing
Media
 Binding factors (albumin, transferrin)
 Increase viscosity of medium
 Protease inhibitor
 Buffer
 Minerals (Na, K, Fe, Zn, and Cu etc)
Disadvantages of
serum
 Inhibit growth of some cell type
 Cytotoxic constituents (polyamine
oxidase)
 Variation in serum quality
 May add growth factors
 Interfere downstream processing
 Lower supply
Serum Free Media
 Analytical approach
 Synthetic approach
 Limiting factor approach

 Advantages of serum free media

 Reproducibility
 Easier Downstream processing
 Avoided toxic effects
 No degradation of sensitive proteins
 Lower cell differentiation
Disadvantage Of Cell
Serum Free Media
 Specific to one cell type
 Media formulation not known commercially
 pH, temp control necessary
 Decrease growth rate and cell density
 Cells become fragile
Defined Media

EMEM, DME, Ham,s F12, CMRL 1066, RPMI

1640, Iscove,s modified Dulbecco,s medium
 Different culture Media
used
Cell type Medium Supplemented with

Permanent cell line MEM. DME, McCDoy,5a, Serum or protein

RPMI1640

Permanent cell line F10, F12,DME Purified proteins and

hormones

Permanent cell line in CMRL 1066, MCDB 411,

monolayer DME, IMDM

Clonal Growth of F12, MCDB 301 etc

permanent cell lines
Chemically Defined
Media
 Contain contamination free ultra pure
inorganic and organic constituents and
may contain pure protein additives like
insulin growth factors etc (produced in
bacteria and yeast)
Protein free Media
Non protein constituents
Assays for
development of best
medium
 Long term
 Clonal growth assay, Cell growth curve
analysis

 Short term
 Thymidine assay
Keep smiling

See ya