Question

Answer the question below: B) Cell Culture/ culture medium: What are the keys to successfully growing cells in culture? What special precautions must be taken? How attentive to the cells must you be? (http://userpages.umbc.edu/~jwolf/method5.htm)

Answer 1

The keys to successfully growing cells in culture are:-

1. The cell culture medium must be having the appropriate energy source to mainting cell cycle of the cell to be cultured. Which means it should have a balance of nitogen source i.e amino acid, carbon source i.e glucose, inorganic and organic salts , vitamines growth factors, hormones and attachment factors specific for the cell to be cultured.

2. The pH and osmolality must have been maintained.

3.Correct temperature for the growth of the cell must have been maintained in the incubator during cell culture.

4. Proper aeration for O₂ or CO₂ or anaerobic condition should have been maintained throughout the cell growth.

5. The cell culture must have been prepared aseptically and maintained in an aseptic condition so as to nullify or decrease the contamination risk.

We should be very attentive to the cells as threre can be a risk of contamination so at a regular interval microscopic check and cell count should be done so that the cells are maintained in exponential phase and in a interval of 1 to 2 days fresh medium should be added as in growing cells there is rapid deplition of medium. However for continous cell lines in conventional medium,two medium changes will maintain exponential cell growth.Since the cells rapidly utilize the nutrients and release metabolites into the medium, resulting in a lowering of pH.The change of medium will also help in maintaing pH of the cell culture.We should monitor the temperature and CO₂ level of incubator which may fluctuate due voltage.