SERUM CONTAINING AND SERUM FREE MEDIA

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Presentation transcript:

SERUM CONTAINING AND SERUM FREE MEDIA

INDEX Intoduction to culture media. Components of serum. Advantages and disadvantages of serum media. Advantages and disadvantages of serum free media.

*extract from spleen , liver CULTURE MEDIA = defined as growth media suitable for the in- vitro cell culturing of human or animal cells and the composition of components are known. CULTURE MEDIA NATURAL 1.BIOLOGICAL FLUIDS * Serum media *amniotic fluid 2.TISSUE EXTRACTS * Chick embryo extract *extract from spleen , liver ARTIFICIAL SERUM FREE MEDIA

COMPONENTS OF SERUM PROTEINS ( ATTACHEMENT AND CARRIER ) ATTACHEMENT = fibronectin and α2 microglobulin . Promotes cell – substrate interaction. CARRIER = albumin and transferrin. albumin carry lipids and vitamins and transferrin carry iron in bioavailable forms. 2. GROWTH FACTORS PDGF , EGF, IGF, FGF , NGF. PDGF has mitogenic activity . Means induce mitosis and help in cell proliferation. 3. GROWTH HORMONE Eg- insulin . Increase uptake of glucose and amino acid. 4. MINERALS Na , Ca 2+ , Mg2+ , Zn2+ act as co-factor & helps in cell metabolic processes. 5. LIPIDS oleic acid , linnolenic acid.

ADVANTAGES OF SERUM complete media as it contains all components require for cell growth. increases viscosity. lesser shear forces arise and viability of cell is more. act as a buffer. no need to add a buffering agent. contains attachments proteins. E.g- fibronectin and no need of charged substrate. Contains minerals Na+ , K + , Zn2+ it contains hormones . E.g insulin. Imp. For cell growth.

DISADVANTAGES OF SERUM Inhibit growth of certain cells. For example- epidermal keratinocytes Contain cytotoxic components. eg- FCS (f0etal calf serum) contains polyamine oxidase that converts polyamines into polyamino aldehydes which are cytotoxic to the cell. Batch to batch variation in serum quality. the experimental results are affected. Growth factor present in the serum may be inadequate and need supplementation. Interfere downstream processing. Increasing expense.

SERUM FREE MEDIA devoid of serum but it contains 10-20% of serum. Examples of serum free media; RPMI = ROSEWELL PARK MEMORIAL INSTITUTE DMEM = DULBECCOS MODIFIED EAGLE’S MEDIA IMDM= ISCOVE’S MODIFIED DULBECCO’S MEDIA immediate survival of the cell HBSS (HANKS BALANCED SALT SOLUTION) and HEPES BUFFER is used. For washing the cell phosphate buffer saline is used.

ADVANTAGES OF SERUM FREE MEDIA Improved and reproducible results because there is no change in quality from batch to batch. Downstream processing is easier. Bioassays are free from cytotoxic components of the serum. No danger of degradation of cellular proteins by the serum proteases. Toxic effects of serum are avoided. Elimination of risk of contaminants . Eg viruses , mycoplasma. Etc.

DISADVANTAGES OF SERUM FREE MEDIA Specific for specific cell type .need to formulate every time for the new experiment. serum free formulations are not available. control of pH and viscosity is required .For maintaining pH hepes buffer is added and to increase the viscosity CMC (CARBOXY METHYL CELLULOSE ) Growth rate and maximum cell density achieved is lower than that of serum containing media. Charged substrate is needed.

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