Abstract
An in vitro system for liver organogenesis from murine embryonic stem (ES) cells has been recently established. This system is expected to be applied to the development of a new drug metabolism assay system that uses ES cells as a substitute for animal experiments. The objective of this study was to elucidate the drug metabolism profiles of the murine ES cell-derived hepatic tissue system compared with those of primary cultures of murine adult and fetal hepatocytes. The expression of the genes of the cytochrome P450 (P450) family, such as Cyp2a5, Cyp2b10, Cyp2c29, Cyp2d9, Cyp3a11, and Cyp7a1, was observed in the murine ES cell-derived hepatic tissue system at 16 days and 18 days after plating (A16 and A18). To investigate the activities of these P450 family enzymes in the murine ES cell-derived hepatic tissue system at A16 and A18, testosterone metabolism in this system was analyzed. Testosterone was hydroxylated to 6β-hydroxytestosterone (6β-OHT), 16α-OHT, 2α-OHT, and 2β-OHT in this system, and was not hydroxylated to 15α-OHT, 7α-OHT, and 16β-OHT. This metabolism profile was similar to that of fetal hepatocytes and different from that of adult hepatocytes. Furthermore, pretreatment with phenobarbital resulted in a 2.5- and 2.6-fold increase in the production of 6β-OHT and 16β-OHT. Thus, evidence for drug metabolic activities in relation to P450s has been demonstrated in this system. These results in this system would be a stepping stone of the research on the development and differentiation to adult liver.
Footnotes
-
This study was supported by grants from the Ministry of Education, Sports, Science and Technology of Japan (Tokyo, Japan) (15700314; 13470150, Grantin-Aid for 21st Century COE program), and the Hokuto Foundation of Bioscience (Nagano, Japan).
-
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
-
doi:10.1124/dmd.105.007674.
-
ABBREVIATIONS: ES, embryonic stem; RT-PCR, reverse transcription-polymerase chain reaction; OHT, hydroxytestosterone; EB, embryoid body; HPLC, high performance liquid chromatography; PB, phenobarbital.
- Received October 2, 2005.
- Accepted January 6, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
DMD articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|