Animal Cell Culture Methods by Jennie P. Mather and David Barnes (Eds.)

By Jennie P. Mather and David Barnes (Eds.)

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This process should be done using the desired end point to screen. For example, if a medium is to be optimized to achieve maximal recombinant protein secretion, then the screen should be done using the protein titer as the end point assayed. If growth is to be optimized, then the cell number is the end point. , 1990). Cells will tolerate a broad range of concentrations for some medium components but have a very narrow optimal concentration range for others. , serum, growth factor mix). For optimizing hormone and growth factor additions for serum-free culture, see Barnes and Sat0 (1980).

When weighing out these reagents, a disposable tongue depressor and weigh boat should be used. This avoids contaminating these reagents with other, potentially toxic, chemicals that may be in use in the laboratory. The major component of the medium is water. Water purity is very important for good quality medium. Usually, water quality is more critical when cells are grown in serum-free medium than when the same cells are grown with serumsupplemented medium. However, some cell types are extremely sensitive to poor medium quality, even when serum is used.

Although it varies from cell to cell, and with serum-supplemented or serum-free media, 2 weeks is a safe storage time for serum-free media and 4-8 weeks if serum is added when the medium is prepared. Outdated medium can be used for washing cells or for preparing tissues for primary culture. Clearly, this is not adequate time to allow for commercial preparation and storage, shipping, and further storage of medium in the laboratory. Some prepared liquid media can be frozen. Those that form a precipitate when thawed should not be frozen.

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