By Jennie P. Mather and David Barnes (Eds.)
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Optimizing the nutrient mixture is an important part of this process. The best way to optimize the nutrient mixture is to sequentially perform dose-response curves on each component, select the optimal range for each, and retest each component. This must be done as an iterative process because the ratios of the components, as well as the absolute levels, are critical in optimizing the medium. 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.
It is best to purchase serum that has been sterilely collected as well as an added insurance against viruses or mycoplasma, which can go through some filters. When adding human serum to cultures, the entire culture and all waste should be treated as a biohazard. Human sera should be collected from known donors or blood banks that test for the most common viruses, such as HIV and hepatitis. VIII. Testing Media and Components and Quality Control: “It’s in the Water” As stated previously, it is best to prepare medium from commercially available powdered nutrient mixtures.
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. , TR-1, a capillary endothelial-derived cell line) cannot be grown at all in serum-free medium made with poor quality water, although they will grow at a decreased rate if this medium is supplemented with serum.