Cell culture media can be natural or artificial. Natural media include coagula/clots, biological fluids, and tissue extracts. Artificial (synthetic) media include serum-containing, serum-free, and chemically defined media. Serum-containing media supplement defined media like Eagle Minimum Essential Medium with 5-20% serum, which provides nutrients, hormones, growth factors, and other important components to support cell growth. However, serum can also inhibit some cell types and contain cytotoxic constituents. Serum-free media avoid these disadvantages but may be specific to cell types and require close control of conditions. Chemically defined media use ultrapure inorganic and organic components and defined protein additives produced through genetic engineering.
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A Culture media is a nutrient solution containing all
essential organic and inorganic ingredients required for
the growth of animal cell.
Various types of media used for tissue culture may be
grouped into two broad categories.
1. Natural media and
2. Artificial media
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NATURAL MEDIA
These media consist of naturally occurring
biological fluids and these are the following three
types
a. Coagula/Clots
b. Biological fluids
c. Tissue extracts
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a. Coagula/Clots:
Most commonly used clots are plasma clots,
which are being used since long time.
Plasma is now commercially available either in
liquid or lyophilized state.
It may be prepared in the laboratory, usually from
blood of male fowl, but clotting must be avoided
during the preparation.
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b. Biological fluids:
Of the various biological fluids used as Culture
media e.g. Amniotic fluid, Ascitic fluid, Pleural
fluid, Aqueous fluid from eye, insect hemolymph,
Serum etc.
Serum is the most widely used. It may be
obtained from human blood, placental cord
blood, horse blood, calf blood.
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c. Tissue extracts:
Chick embryo extracts is the most
commonly used tissue extracts
but bovine embryo extracts are also used
other tissue extracts that have been used
are Spleen, liver, bone marrow, leucocytes
etc. extracts.
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ARTIFICIAL MEDIA OR SYNTHETIC MEDIA
Various artificial media may be
grouped into following types.
Serum containing media
Serum free media and
Chemically defined media
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a. Serum containing media:
Various defined media, e.g. Eagle Minimum Essential
Medium (EMEM) etc., When supplemented with 5-20%
serum becomes good nutrient medium for culture of most
types of cells. Serum serves the following major functions.
It provides the basic nutrients for cell. The nutrients are
present both in the solution and are bound to the
proteins.
It provides hormones like insulin, which is essential for
growth of nearly all cells in culture.
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It contains several growth factors like Platelet –
derived growth factor, transforming growth factor,
epidermal growth factor, fibroblast growth factor,
endothelial growth factor etc.
A major role of serum is to supply proteins. Like
fibronectin, to promote attachment of cells to the
substrate.
It provides several binding proteins like Albumin,
Transferrin etc. which carry other molecules into the
cell.
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It increases the viscosity of medium and
thereby, protects cells from mechanical
damages.
The serum also provides several minerals
like Na+, k+, Fe+, Zn+, Cu++, etc.
It also acts as a buffer.
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Disadvantages of using serum in the culture media
Serum may inhibit growth of some cell types such
as epidermal keratinocytes.
Serum may contain potentially cytotoxic
constituents. Ex: feotal calf serum contain the
enzyme polyamine oxidase, which converts
polyamines like spermidine and spermine into
cytotoxic poly amino aldehydes.
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b. Serum free media:
In view of the disadvantages due to serum,
extensive investigations have been made to develop
serum free formulations of culture media.
Improved reproducibility of results from different
laboratories.
Easier downstream processing of products from
cultured cells.
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Toxic effects of serum are avoided.
There is no danger of degradation of serum
proteins.
Permit selective culture of differentiated
cells and produce cell types from the
heterogenous cultures.
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Disadvantages:
Most serum free media are specific to one cell type.
Therefore, different media may be required for different
cell lines.
Reliable serum-free media preparation, for most of the
media formulations are not available commercially.
A greater control of pH, temperature etc. is necessary as
compared with serum containing media.
Growth rate and the maximum cell density attained are
lower than those with serum containing media.
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c. Chemically defined media:
These media contain contamination-
free ultrapure inorganic and organic
constituents, besides pure protein additives
like insulin, epidermal growth factor etc.
that have been produced in bacteria or
yeast by genetic engineering methods.
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