By Jennie P. Mather and David Barnes (Eds.)
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Extra resources for Animal Cell Culture Methods
Test all cell lines in the laboratory for mycoplasma and other potential contaminants. If they are all contaminant free and only one cell line seems to be having trouble, thaw out a vial of cells from an earlierfreeze of that line. 7. If any of these questions have turned up a suspicious reagent or supply, test this first. Make up medium using a different lot of medium powder, 2. Choosing the Appropriate Medium and Culture System 27 serum, and so on (or open another lot of tissue culture dishes) and test by comparing the newly made medium to the presumptive “bad” lot of medium.
Sometimes an increased concentration of vitamins can be useful. Vitamin mixtures are commercially available and can be added as such. Some vitamins, such as vitamin E (or a-tocopherol), A (retinol or retinoic acid), or C , are not added to media mixtures because of their instability, but may be important for some cells to survive or function in vitro. To use vitamin E or A, make a 1000-fold stock solution in absolute ethanol (not benzene distilled) and dilute in an aliquot of medium immediately before use.
This avoids any possibility of detergent residue getting into the medium. Bottles of sodium bicarbonate and any other reagents that are used in medium should be used only for medium. 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.
Animal Cell Culture Methods by Jennie P. Mather and David Barnes (Eds.)