Media For Cell Culture

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

Permanent cell line in monolayer

CMRL 1066, MCDB 411, DME, IMDM

Clonal Growth of permanent cell lines

F12, MCDB 301 etc

Purified proteins and hormones

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

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