The use of primary rat hepatocytes to achieve metabolic activation of promutagens in the Chinese hamster ovary/hypoxantine-guanine phosphoribosyl transferase mutational assay
A method is described in which primary rat hepatocytes have been cocultured with chinese hamster ovary (CHO) cells to provide metabolic activation of promutgens in the Chinese hamster ovary/hypoxanthine-guanine phosphoribosyl transferase (CHO/HGPRT) mutational assay. Single cell hepatocyte suspensions were prepared from male Fisher-344 rats using the in situ collagenase perfusion technique. Hepatocytes were allowed to attach for 1.5 hours in tissue culture dishes containing an approximately equal number of CHO cells in log growth. The cocultures were exposed to promutagens for up to 20 hours in serum-free medium. The survival and 6-thioguanine-resistant fraction of treated CHO cells were then determined as in the standard CHO/HGPRT assay. Aflatoxin B/sub 1/ (AFB/sub 1/) 7,12-dimethylbenz(a)anthracene (DMBA) and benzo(a)pyrene (B(a)P) were found to produce increases in the mutant fractions of treated CHO cells as a function of concentration. The time required for optimum expression of the mutant phenotype following exposure to DMBA and AFB/sub 1/ was approximately 8 days. Primary cell-mediated mutagenesis may be useful in elucidating methobolic pathways important in the production and detoxification of genotoxic products in vivo.
- Research Organization:
- Chemical Industry Inst. of Toxicology, Research Triangle Park, NC
- OSTI ID:
- 5336825
- Journal Information:
- Environ. Mutagen.; (United States), Vol. 4:1
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
MUTAGENS
METABOLIC ACTIVATION
AFLATOXIN
BENZANTHRACENE
BENZOPYRENE
ENZYME ACTIVITY
HAMSTERS
HOMOGENATES
LIVER
LIVER CELLS
MICROSOMES
MUTAGEN SCREENING
MUTAGENESIS
OVARIES
PHENOTYPE
RATS
ANIMAL CELLS
ANIMALS
AROMATICS
BODY
CELL CONSTITUENTS
CONDENSED AROMATICS
DIGESTIVE SYSTEM
FEMALE GENITALS
GLANDS
GONADS
HYDROCARBONS
MAMMALS
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
RODENTS
SCREENING
SOMATIC CELLS
VERTEBRATES
560301* - Chemicals Metabolism & Toxicology- Cells- (-1987)