The use of primary rat hepatocytes to achieve metabolic activation of promutagens in the Chinese hamster ovary/hypoxantine-guanine phosphoribosyl transferase mutational assay

Journal Article · Fri Jan 01 04:00:00 EST 1982 · Environ. Mutagen.; (United States)

DOI:https://doi.org/10.1002/em.2860040108· OSTI ID:5336825

Bermudez, E; Couch, D B; Tillery, D

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), Journal Name: Environ. Mutagen.; (United States) Vol. 4:1; ISSN ENMUD
• Country of Publication: United States
• Language: English

Related Subjects

560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AFLATOXIN
ANIMAL CELLS
ANIMALS
AROMATICS
BENZANTHRACENE
BENZOPYRENE
BODY
CELL CONSTITUENTS
CONDENSED AROMATICS
DIGESTIVE SYSTEM
ENZYME ACTIVITY
FEMALE GENITALS
GLANDS
GONADS
HAMSTERS
HOMOGENATES
HYDROCARBONS
LIVER
LIVER CELLS
MAMMALS
METABOLIC ACTIVATION
MICROSOMES
MUTAGEN SCREENING
MUTAGENESIS
MUTAGENS
ORGANIC COMPOUNDS
ORGANOIDS
ORGANS
OVARIES
PHENOTYPE
RATS
RODENTS
SCREENING
SOMATIC CELLS
VERTEBRATES