Sister chromatid exchange studies in human fibroblast-rat hepatocyte co-cultures: a new in vitro system to study SCEs.

To approximate better the metabolic reactions that take place in vivo yet maintain the simplicity and reproducibility of in vitro systems, we have developed a co-culture system making use of freshly isolated rat hepatocytes and confluent human fibroblasts for the study of SCE induction by genotoxic agents. Hepatocytes were obtained from male rats by reverse collagenase perfusion and plated over low-passage male human fibroblasts. Preliminary studies demonstrated that although the number of hepatocytes plated was not critical, the best attachment to, and coverage of, the fibroblasts occurred when between 10 x 10(6) and 20 x 10(6) hepatocytes were plated/100 mm tissue culture dish. Results with the promutagen cyclophosphamide showed that the hepatocytes could metabolize the compound and deliver active moieties to the fibroblasts resulting in a linear dose-dependent increase in SCE frequencies. Control fibroblast cultures lacking hepatocytes displayed no increase in SEC frequencies. Control fibroblast cultures lacking hepatocytes displayed no increase in SEC frequencies following cyclophosphamide administration.