A practical cytogenetic protocol for in vitro cytotoxicity and genotoxicity testing

In vitro cytogenetics has been established as a valid method for evaluating the genotoxic potential of chemical agents. Armstrong et al have described a simple, quantitative approach to in vitro cytotoxicity and genotoxicity testing by using Chinese hamster ovary (CHO) cells. This approach can also be sensitive and repeatable in an inter-laboratory setting, a prerequisite for routine testing of compounds suspected of having genotoxic properties. In the present study, cytotoxicity was evaluated by the parameter of mitotic index (MI). Genotoxicity is measured by the chromosome aberration (Abs) assay as described by Armstrong et al using CHO cells. The basic analytic principles proposed were extended to include human lymphocytes. Sister chromatid exchange (SCE) analysis was used to establish an additional endpoint. Mitomycin C (MMC), an established clastogen, was used as the model compound for protocol validation. Dose response curves for MI and Abs in CHO cells were found to be consistent with those reported by Armstrong et al. Results from our extended study on lymphocytes and using SCE analysis were analogous. Our experience is that this standardized approach is indeed sensitive and reliable and can serve as a basis for an inter-laboratory testing program.