Mutagenicity testing in mammalian cells: multiple drug-resistance markers

Technical Report · Wed Aug 01 00:00:00 EDT 1979

OSTI ID:5511183

Carver, J H; Adair, G M; Wandres, D L

In this study, we describe the derivation of a CHO cell line that is heterozygous for both the adenine phosphoribosyltransferase (aprt) and thymidine kinase (tk) loci. This subline allows single-step selection of autosomal recessive AA/sup r/ or FUdR/sup r/ mutant phenotypes as well as the more commonly used OUA/sup R/ or TG/sup r/ genetic markers. Biochemical and genetic characterization of the heterozygous cell line, and dose response data for mutation induction at these four genetic loci by the direct-acting mutagen, ethyl methanesulfonate (EMS), are presented. We also establish optimal conditions for the phenotypic expression and selection of AA/sup r/ and FUdR/sup r/ mutants of CHO cells and present biochemical validation of the mutant phenotypes. Optimal drug concentrations, cell plating densities, and expression time requirements are determined for all four drug-resistance markers. Mutation data are reported for direct mutagens (EMS, MNNG, NQO) and promutagens requiring metabolic activation (DMN, BP). Finally, we discuss the role and expected use of the multiple-marker mutagenesis assay to yield increased sensitivity and reliability in detecting genetic damage induced by complex pollutant mixtures of environmental concern.

• Research Organization: California Univ., Livermore (USA). Lawrence Livermore Lab.
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 5511183
• Report Number(s): UCID-18599; EPA-600/7-79-173
• 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.
ANIMAL CELLS
ANIMALS
CELL CULTURES
EVALUATION
HAMSTERS
IN VITRO
MAMMALS
MUTAGEN SCREENING
PERFORMANCE TESTING
POLLUTION
RODENTS
SCREENING
TESTING
VERTEBRATES