RT Journal Article
SR Electronic
T1 Differential Maintenance of Cytochrome P450 Enzymes in Cultured Precision-Cut Human Liver Slices
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 1202
OP 1209
VO 28
IS 10
A1 Renwick, Anthony B.
A1 Watts, Patricia S.
A1 Edwards, Robert J.
A1 Barton, Paula T.
A1 Guyonnet, Isabelle
A1 Price, Roger J.
A1 Tredger, J. Michael
A1 Pelkonen, Olavi
A1 Boobis, Alan R.
A1 Lake, Brian G.
YR 2000
UL http://dmd.aspetjournals.org/content/28/10/1202.abstract
AB 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 supplemented RPMI 1640 medium. The relative apoprotein levels of 11 CYP enzymes were determined using a panel of antipeptide antibodies. In addition, 7-ethoxyresorufin O-deethylase, tolbutamide methylhydroxylase, debrisoquine 4-hydroxylase, and testosterone 6β-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 relatively stable (half-lives between 70 and 104 h), compared with CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C19, CYP2E1, and CYP3A5, which were relatively unstable (half-lives between 23 and 36 h). Enzyme activities decreased at rates similar to those of their corresponding apoproteins. There was also a large difference in the stability of individual CYP enzymes from different liver donors, particularly for the most rapidly declining CYP enzymes. Similar losses of CYP enzymes were found when human liver slices were cultured in supplemented Williams' medium E for 72 h, except that CYP2E1 apoprotein levels were better maintained. Because of the variable decreases of CYP enzymes, xenobiotic metabolism studies are best performed with freshly cut rather than cultured human liver slices. The American Society for Pharmacology and Experimental Therapeutics