Induction of Human CYP1A2 and CYP3A4 in Primary Culture of Hepatocytes from Chimeric Mice with Humanized Liver

• Published in: Drug metabolism and pharmacokinetics Vol. 20; no. 2; pp. 121 - 126
• Main Authors: Nishimura, Masuhiro, Yokoi, Tsuyoshi, Tateno, Chise, Kataoka, Miho, Takahashi, Eiji, Horie, Toru, Yoshizato, Katsutoshi, Naito, Shinsaku
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 2005
• Subjects: Adolescent; Animals; Cells, Cultured; Chimera; chimeric mouse liver; Cryopreservation; CYP1A2; CYP3A4; Cytochrome P-450 CYP1A2 - biosynthesis; Cytochrome P-450 CYP3A; Cytochrome P-450 Enzyme System - biosynthesis; Enzyme Induction; Hepatocytes - enzymology; human hepatocytes; Humans; induction; Male; Mice; real-time RT-PCR; Reverse Transcriptase Polymerase Chain Reaction; chimeric mouse liver; human hepatocytes; induction; CYP3A4; real-time RT-PCR; CYP1A2
• ISSN: 1347-4367; 1880-0920
• DOI: 10.2133/dmpk.20.121

Chimeric mice with near-completely humanized liver were constructed by transplantating hepatocytes from a Japanese and Caucasian donor. In the present study, we investigated the induction of human CYP1A2 and CYP3A4 mRNA in a primary culture of the cryopreserved chimeric mouse hepatocytes. β-naphthoflavone (β-NF) and rifampicin (Rif) were used as typical cytochrome P450 (CYP) inducers for CYP1A2 and CYP3A4, respectively. Analysis was performed by the real-time reverse-transcription polymerase chain reaction method. CYP1A2 mRNA in the primary culture of chimeric mouse hepatocytes in mice No. 1, 2, and 3 was significantly increased 3.8-, 6.3-, and 3.3-fold by 5 μΜ β-NF exposure, respectively, compared with the 0.1% DMSO treated control (p<0.01). CYP3A4 mRNA in the primary culture of chimeric mouse hepatocytes in mice No. 1, 2, and 3 was significantly increased 8.4-fold (p<0.001), 2.2-fold (p<0.01), and 2.3-fold (p<0.05) by 50 μΜ Rif exposure, respectively, compared with the 0.1% DMSO treated control. The present study demonstrated that a primary culture of cryopreserved hepatocytes from chimeric mice with humanized liver could be used for evaluating the induction of drug metabolizing enzymes in human. This in vitro method may be a useful method for screening the induction potency of new drug candidates on drug metabolizing enzymes.