Cytochrome P450 activities in pure and co-cultured rat hepatocytes. Effects of model inducers.

The stability and inducibility of several P450 activities (namely, P450 1A1; 2A1, 2B1/2, 2C11, and 3A1) were studied in rat hepatocytes co-cultured with the MS epithelial cell line derived from monkey kidney. The results revealed that these monooxygenase activities were systematically higher in co-cultures than in conventional hepatocyte cultures. Pure cultures showed a rapid loss of monooxygenase activities, which were undetectable after 5 days. In contrast, all isozymes assayed were measurable in co-cultured hepatocytes on Day 7 (about 15 to 40% of the initial activities of Day 0 of culture). The beneficial effects of the co-culture system seemed to be more selective for certain cytochrome P450 isoforms, with P450 1A1 and 3A1 being the best stabilized isozymes after 1 wk. A clear response to inducers was observed in co-cultures, each isozyme showing a different induction pattern. 3-Methylcholanthrene produced a strong increase in P450 1A1 (7-ethoxyresorufin O-deethylase) activity and a low increase in P450 2A1 (testosterone 7 alpha-hydroxylation), whereas no changes were observed in the other activities. Phenobarbital treatment resulted in increases in P450 2B1/2 (7-pentoxyresorufin O-depentylase and 16 alpha- and 16 beta-hydroxylation of testosterone) activities, while minor effects were observed on P450 3A1 (testosterone 6 beta-hydroxylation) activity. Dexamethasone markedly increased P450 3A1 (testosterone 6 beta- and 15 beta-hydroxylation) activity and, to a lesser extent, P450 2B1/2 (16 beta-hydroxylation).