Differential maintenance of cytochrome P450 enzymes in cultured precision-cut human liver slices

The maintenance of the major hepatic cytochrome P450 (CYP) enzymes has been studied in precision-cut human liver slices cultured for up to 72 h in sup plemented RPMI 1640 medium. The relative apoprotein levels of 11 CYP enzyme s were determined using a panel of antipeptide antibodies. In addition, 7-e thoxyresorufin O-deethylase, tolbutamide methylhydroxylase, debrisoquine 4- hydroxylase, and testosterone 6 beta-hydroxylase activities were determined as enzymatic markers for CYP1A2, CYP2C9, CYP2D6, and CYP3A4, respectively. There was a large variation in the rate of decline of different CYP levels with time in culture. Based on the rate of decrease, CYP enzymes could be separated into two groups, with CYP2C9, CYP2D6, CYP3A4, and CYP4A11 being r elatively stable (half-lives between 70 and 104 h), compared with CYP1A2, C YP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2E1, and CYP3A5, which were relatively u nstable (half-lives between 23 and 36 h). Enzyme activities decreased at ra tes similar to those of their corresponding apoproteins. There was also a l arge difference in the stability of individual CYP enzymes from different l iver donors, particularly for the most rapidly declining CYP enzymes. Simil ar losses of CYP enzymes were found when human liver slices were cultured i n supplemented Williams' medium E for 72 h, except that CYP2E1 apoprotein l evels were better maintained. Because of the variable decreases of CYP enzy mes, xenobiotic metabolism studies are best performed with freshly cut rath er than cultured human liver slices.