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INDEX

Intoduction to culture media.

Components of serum.

Advantages and disadvantages of serum media.

Advantages and disadvantages of serum free media.

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
1.
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+
,
5. LIPIDS
oleic acid , linnolenic acid.
Zn2+ act as co-factor & helps in cell metabolic processes.
ADVANTAGES OF SERUM
1.
complete media as it contains all components require for cell growth.
2.
increases viscosity. lesser shear forces arise and viability of cell is more.
3.
act as a buffer. no need to add a buffering agent.
4.
contains attachments proteins. E.g- fibronectin and no need of charged
substrate.
5.
Contains minerals Na+ , K + , Zn2+
6.
it contains hormones . E.g insulin. Imp. For cell growth.
DISADVANTAGES OF SERUM
1.
Inhibit growth of certain cells. For example- epidermal
keratinocytes
2.
Contain cytotoxic components. eg- FCS (f0etal calf serum)
contains polyamine oxidase that converts polyamines into
polyamino aldehydes which are cytotoxic to the cell.
3.
Batch to batch variation in serum quality. the experimental
results are affected.
4.
Growth factor present in the serum may be inadequate and
need supplementation.
5.
Interfere downstream processing. Increasing expense.
SERUM FREE MEDIA

devoid of serum but it contains 10-20% of serum.

Examples of serum free media;
1.
RPMI = ROSEWELL PARK MEMORIAL INSTITUTE
2.
DMEM = DULBECCOS MODIFIED EAGLE’S MEDIA
3.
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
1.
Improved and reproducible results because there is no change
in quality from batch to batch.
2.
Downstream processing is easier.
3.
Bioassays are free from cytotoxic components of the serum.
4.
No danger of degradation of cellular proteins by the serum
proteases.
5.
Toxic effects of serum are avoided.
6.
Elimination of risk of contaminants . Eg viruses , mycoplasma.
Etc.
DISADVANTAGES OF SERUM FREE MEDIA
1.
2.
Specific for specific cell type .need to formulate every time
for the new experiment.
serum free formulations are not available.
3.
control of pH and viscosity is required .For maintaining pH
hepes buffer is added and to increase the viscosity CMC
(CARBOXY METHYL CELLULOSE )
4.
Growth rate and maximum cell density achieved is lower than
that of serum containing media.
5.
Charged substrate is needed.