Chinese hamster ovary cells cultured in low concentrations of fetal bovine serum: cloning efficiency, growth in suspension, and selection of drug-resistant mutant phenotypes
Journal Article
·
· In Vitro; (United States)
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% FBS plus BSA; observed mutant frequencies induced by dimethylnitrosamine or benzo(a)pyrene seemed to be decreased at lower survival levels.
- Research Organization:
- Univ. of California, Livermore
- DOE Contract Number:
- W-7405-ENG-48
- OSTI ID:
- 6355124
- Journal Information:
- In Vitro; (United States), Journal Name: In Vitro; (United States) Vol. 19:9; ISSN ITCSA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550300* -- Cytology
550400 -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BODY
CELL CULTURES
CLONING
COMPARATIVE EVALUATIONS
CULTURE MEDIA
DRUGS
EFFICIENCY
FEMALE GENITALS
GONADS
GROWTH
HAMSTERS
IMMUNE REACTIONS
MAMMALS
MUTAGEN SCREENING
MUTATIONS
ORGANS
OVARIES
PHENOTYPE
RODENTS
SCREENING
SPONTANEOUS MUTATIONS
VERTEBRATES
550400 -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BODY
CELL CULTURES
CLONING
COMPARATIVE EVALUATIONS
CULTURE MEDIA
DRUGS
EFFICIENCY
FEMALE GENITALS
GONADS
GROWTH
HAMSTERS
IMMUNE REACTIONS
MAMMALS
MUTAGEN SCREENING
MUTATIONS
ORGANS
OVARIES
PHENOTYPE
RODENTS
SCREENING
SPONTANEOUS MUTATIONS
VERTEBRATES