Abstract
Publisher Summary This chapter describes Chinese hamster ovary (CHO) cells that use for genetic analysis in tissue culture. These cells have been used for the isolation of mutants affecting intermediary metabolism; DNA, RNA, and protein synthesis; membrane functions; and several more complex forms of cell behavior such as cell growth and endocytosis. A recent compilation of CHO mutants lists more than 80 classes of mutants that are isolated using this cell line. The methodologies described in this chapter are developed for work with the CHO-S sublines but most of the methods, with the exception of suspension culture, may be used for CHO-K1 cell lines. CHO cells are proline auxotrophs, unlike most other cultured cell lines, and require medium containing this nutrient, such as Ham's F12 or a-modified Eagle's medium (a-MEM) without ribonucleoside or deoxyribonucleosides, both of which are commercially available. These rich media must in addition contain other limiting nutrients for CHO cells, as they support more rapid growth of CHO lines than is possible in MEM alone supplemented with proline.
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