In vitro testing of an indirect mutagen (cyclophosphamide) with human leukocyte cultures: activation with liver microsomes and use of a dialysis bag.

A system of cell-mediated mutagenesis is described for the study of compounds which require metabolic activation to exert their cytotoxic and mutagenic effects. This system combines BHK21 cells for metabolism of the compounds and V79 cells as targets for mutagenesis. Using the two polycyclic hydrocarbon carcinogenesis benzo(a)pyrene and 7-methylbenz(a)anthracene we have shown that the hydrocarbon-DNA reaction which accompanies mutagenesis in the target cell is indistinguishable from that reported to occur in vivo and in primary cell cultures. Our results also support the view that a diol epoxide metabolite is responsible for the biological activity of benzo(a)-pyrene.The application of cell-mediated mutagenesis to the routine testing of suspect environmental chemicals for biological activity is discussed.