Professional Documents
Culture Documents
u The nutrient media used for culture of animal cells and tissues
must be able to support their survival as well as growth, i.e.,
must provide nutritional, hormonal and stromal factors.
1. Most serum free media are specific to one cell type. Therefore,
different media may be required for different cell lines.
2. Reliable serum free preparations, for most of the media
formulations are not available commercially. This necessitates
time consuming task of preparing the desired formulations in the
laboratory.
3. A greater control of pH, temperature, etc. is necessary as
compared to that with serum containing media.
4. Growth rate and the maximum cell density attained are lower
than those with serum containing media.
5. Cells tend to become fragile during prolonged agitated cultures
unless biopolymers or synthetic polymers are added.
u Several defined media have been evolved from the Eagle's
minimal essential medium (MEM),
u e.g., Dulbecco's enriched modification (DME),
u Ham's F12, :,pMR1O66, RPMIl640,
u McCoy's 5A and
u Iscove's modified Dulbecco¶s' (IMDM); all are commercially
available.
u Often a 1: 1 mixture of DME and F12 is used as a serum free
formulation.
u Chem lly e ne Me D
These media contain contamination free ultra pure inorganic and
organic constituents, and may contain pure protein additives, like
insulin, epidermal growth factor, etc. that have been produced in
bacteria or yeast by genetic engineering.
u Ñ ote n F ee Me -
In contrast, protein free media do not contain any protein; they
only contain non-protein constituents necessary for culture of the
cells. The formulations MEM, DME, RPM-1640, etc. are protein
free; where required, protein supplementation is provided.
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|ntges sntges
1. Provides basic nutrient for cells, nutrients being present in the Inhibits growth of some cell types,
solution and also bound to proteins. e.g. epidermal keratinocytes.
2. Provides several hormones e.g. insulin, testosterone, prostaglandin, May contain some cytotoxic
etc., which are essential for cell growth constituents.
3. Contains several growth factors, e.g. platelet derived growth factor Serum quality varies from batch to
(PDGF). batch which requires costly
and time-consuming tests for
each batch.
4. Supplies proteins, e.g. fibronectin promoting attachment of cells and Some growth factors are inadequate
spreading. for specific cell types.
5. Provides several binding proteins, e.g. albumin and transferrin, which When cell cultures are used for
carry other molecules into the cell. production of biochemicals,
the serum interferes with
downstream processing.
6. Increases viscosity of the medium. thereby protecting cells from Supply of serum is always lower
mechanical damage. than its demand.
7. Protease inhibitors in the serum protect cells from proteolysis.
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