ANIMAL CELL CULTURE MAINTENANCE
Why Animal Cell Culture Maintenance?
As we know cell culture(Primary Cell Culture) or tissue culture is the procedure to grow cells or tissues under totally controlled condition.but cell culture affected by contamination or microbial infection that causes fail to process. for this requires maintenance of cells for optimal cell health. so many ways used to overcome this problem,
Today a variety of ways are available to cell culture incubators, vessels, and antibiotics that allow tightly conditions for culture to avoid contamination and infection. but sometimes require to maintain to culture for a long time for this the best method called cryo-preservation or frizzing of culture by the use of cryoprotectant DMSO(dimethyl sulfoxide) that is a toxic agent. in this process cells preserve at very low temperature and on this cell tolerate to very low metabolic activity.
- laminar air flow hood.
- mechanical pipetter.
- inverted microscope.
- vacuum pump and flask.
- Co2 incubator.
Process- (Cell Culture Protocol)
- use only healthy cells or tissues.
- detach cells with trypsin.
- than resuspend cells to the growth medium.
- adjust concentration about 4–8 x 106 cells/ml.
- than filled to culture in specific vials or ampules.
- then freeze them slowly.
then transfer to vials in gaseous phase mean at liquid nitrogen (-196°C) with liquid nitrogen,
Recovery of preserved cells-
In this process, preserved ampules transfer on a water bath at 36°C. then transfer to cells on appropriate medium and allowing to produced cell monolayer,
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