PT  - JOURNAL ARTICLE
AU  - Roymans, Dirk
AU  - Annaert, Pieter
AU  - Van Houdt, Jos
AU  - Weygers, Adri
AU  - Noukens, Jan
AU  - Sensenhauser, Carlo
AU  - Silva, José
AU  - Van Looveren, Cis
AU  - Hendrickx, Jan
AU  - Mannens, Geert
AU  - Meuldermans, Willem
TI  - EXPRESSION AND INDUCTION POTENTIAL OF CYTOCHROMES P450 IN HUMAN CRYOPRESERVED HEPATOCYTES
AID  - 10.1124/dmd.104.003046
DP  - 2005 Jul 01
TA  - Drug Metabolism and Disposition
PG  - 1004--1016
VI  - 33
IP  - 7
4099  - http://dmd.aspetjournals.org/content/33/7/1004.short
4100  - http://dmd.aspetjournals.org/content/33/7/1004.full
SO  - Drug Metab Dispos2005 Jul 01; 33
AB  - Fresh human hepatocytes are still considered as the “gold standard” to screen in vitro for cytochrome P450 (P450) induction. However, sparse availability of good quality human liver tissue for research purposes and the demand for standardized cell populations, together with the need for proper storage of the cells not immediately required, have resulted in the development of cryopreservation techniques that provide adequate viability and plateability of hepatocytes after thawing. This study aimed at validating cryopreserved human hepatocytes as a model to investigate P450 induction. Cryopreserved cells from four different donors were plated and cultured for 48 h, followed by incubation in the presence of typical P450 inducers. During the experiments, quality of the cultured cells was monitored both physiologically and morphologically. Concomitantly, the activity of CYP1A2, 2B6, 2C9, 2E1, and 3A4 was measured together with their mRNA and protein expression. Determination of CYP1A2, 2B6, 2C9, 2E1, and 3A4 activity in control versus prototypical inducer-treated hepatocytes revealed a maximal significant mean 11.6-, 2.8-, 1.9-, 1.5-, and 9.0-fold induction over their basal expression, respectively. Protein expression analysis of these P450s confirmed these results. Moreover, a mean 44.9-, 3.5-, 3.2-, and 13.8-fold induction of CYP1A2, 2B6, 2C9, and 3A4 mRNA was observed. Our data demonstrate that cryopreserved human hepatocytes are a valuable tool to study the induction of CYP1A2, 2B6, 2C9, 2E1, and 3A4. The American Society for Pharmacology and Experimental Therapeutics