Write a note on routine maintenance of cell lines in vitro.
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Cell culture is the process by which cells are grown under controlled conditions, generally outside their natural environment. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions.
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Mammalian cell culture" refers to the cells of mammals, isolated from specific tissues (i.e., skin, liver, glands, etc.) and further cultivated and reproduced in artificial medium. It becomes a major field in modern biotechnology, especially in the area of human health.
During the cultivation of mammalian cells in vitro (ie outside a living organism), some specific difficulties arise in the extraction of cells from a "safe" tissue. Slow growth rates with doubling times in between 12 and 28 h, low sensitivity, productivity against shear stress due to the lack of a cell wall and complex requirements of growth medium are the challenges in developing techniques for mammalian cell culture. Moreover, many cell lines grow in an adherent manner and a suitable surface for attachment has to be provided for these cells to proliferate. Mammalian cells have originated from the multi-cellular organisms, they still carried the genetic program of inducing their own cell death; a process called "apoptosis" or "programmed cell death". It can limit culture productivity in biotechnological processes. Another important problem is the finite life-span of primary cells, die after several doublings in vitro. This problem was solved by must be sub cultured or passage.
When mammalian cells are cultured in vitro, the investigator is attempting to reproduce physiological environment in order to maintain and analyze the normal responses and functions. The culture medium is an important component of the in vitro environment.
For tissue culture media many different recipes are available and each laboratory must determine which medium best suits their needs. Individual laboratories used commercially prepared medium or self prepared. Commercially available medium are highly sterile and ready-to-use liquid, in a concentrated liquid or powdered form.Instead of providing nutrients for growing cells, medium is generally supplemented with fungicides and antibiotics, or both to inhibit contamination.