title: Characterization of cytochrome P450 expression in murine embryonic stem cell-derived hepatic tissue system.
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 6beta-hydroxytestosterone (6beta-OHT), 16alpha-OHT, 2alpha-OHT, and 2beta-OHT in this system, and was not hydroxylated to 15alpha-OHT, 7alpha-OHT, and 16beta-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 6beta-OHT and 16beta-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.
authors.affiliation: Development Research, R and D, Kissei Pharmaceutical Co., Ltd., Nagano, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Tsutsui
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ogawa
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Inada
authors.forename: Eisuke
authors.initials: E
authors.lastname: Tomioka
authors.forename: Akiko
authors.initials: A
authors.lastname: Kamiyoshi
authors.forename: Satoru
authors.initials: S
authors.lastname: Tanaka
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Kishida
authors.forename: Masahiko
authors.initials: M
authors.lastname: Nishiyama
authors.forename: Makoto
authors.initials: M
authors.lastname: Murakami
authors.forename: Junji
authors.initials: J
authors.lastname: Kuroda
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Hashikura
authors.forename: Shinichi
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Fumiyasu
authors.initials: F
authors.lastname: Satoh
authors.forename: Nobuo
authors.initials: N
authors.lastname: Shibata
authors.forename: Yoh-Ichi
authors.initials: Y
authors.lastname: Tagawa
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004622:Embryo, Mammalian; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D006900:Hydroxylation; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D010634:Phenobarbital / Q000494:pharmacology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D012333:RNA, Messenger / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D013997:Time Factors
chemical_list: D007527:Isoenzymes; D012333:RNA, Messenger; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; D010634:Phenobarbital