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Title: Chinese hamster ovary cells cultured in low concentrations of fetal bovine serum: cloning efficiency, growth in suspension, and selection of drug-resistant mutant phenotypes

Reducing serum concentrations in media provides a potential cost advantage. To determine whether such media could be used for applied mutagenesis assays, cloning efficiency and growth parameters in suspension of Chinese hamster ovary cells cultured were measured in reduced serum with or without additives (1 ..mu..g/ml insulin, 3 x 10/sup -7/ M linoleic acid, 1 x 10/sup -8/ M H/sub 2/SeO/sub 3/) or bovine serum albumin (BSA, 1% wt/vol). With the additives and less than or equal to 0.5% fetal bovine serum (FBS), Ham's F12 medium (without hypoxanthine and thymidine) was more optimal than alpha Eagle's minimum essential medium (MEM) (without ribosides and deoxyribosides) for low density cloning and high density suspension growth. The spontaneous frequency of azaadenine-resistant phenotypes (mutant at the aprt locus) in 1% FBS plus BSA was significantly lower than the frequency observed in 2% FBS plus BSA or 10% DFBS. Frequencies of spontaneous mutants resistant to thioguanine (hgprt locus) or fluorodeoxy-uridine (tk locus) were similar with 10% DFBS, 1% FBS plus BSA, or 2% FBS plus BSA. Compared to alpha MEM with 10% DFBS, frequencies of drug-resistant mutants induced by ethyl methanesulfonate or mitomycin C (MMC) were not significantly lower in alpha MEM with 2% FBSmore » plus BSA; observed mutant frequencies induced by dimethylnitrosamine or benzo(a)pyrene seemed to be decreased at lower survival levels.« less
Authors:
; ;
Publication Date:
OSTI Identifier:
6355124
DOE Contract Number:
W-7405-ENG-48
Resource Type:
Journal Article
Resource Relation:
Journal Name: In Vitro; (United States); Journal Volume: 19:9
Research Org:
Univ. of California, Livermore
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CELL CULTURES; GROWTH; CLONING; EFFICIENCY; CULTURE MEDIA; COMPARATIVE EVALUATIONS; DRUGS; HAMSTERS; IMMUNE REACTIONS; MUTAGEN SCREENING; OVARIES; PHENOTYPE; SPONTANEOUS MUTATIONS; ANIMALS; BODY; FEMALE GENITALS; GONADS; MAMMALS; MUTATIONS; ORGANS; RODENTS; SCREENING; VERTEBRATES 550300* -- Cytology; 550400 -- Genetics