Open AccessCulture density contributes to hepatic functions of fresh human hepatocytes isolated from chimeric mice with humanized livers: Novel, long-term, functional two-dimensional in vitro tool for developing new drugs
Author(s) -
Chihiro Yamasaki,
Yuji Ishida,
Ami Yanagi,
Yasumi Yoshizane,
Yuha Kojima,
Yuko Ogawa,
Yutaka Kageyama,
Yukiko Iwasaki,
Susumu Ishida,
Kazuaki Chayama,
Chise Tateno
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0237809
Subject(s) - pregnane x receptor , in vitro , cytochrome p450 , biology , bone canaliculus , transporter , drug metabolism , multidrug resistance associated protein 2 , in vivo , gene expression , microbiology and biotechnology , pharmacology , atp binding cassette transporter , biochemistry , transcription factor , drug , gene , metabolism , nuclear receptor , genetics , anatomy
Chimeric mice with humanized livers are considered a useful animal model for predicting human (h-) drug metabolism and toxicity. In this study, the characteristics of fresh h-hepatocytes (cFHHs, PXB-cells ® ) isolated from chimeric mice (PXB-mice ® ) were evaluated in vitro to confirm their utility for drug development. cFHHs cultured at high density (2.13 × 10 5 cells/cm 2 ) displayed stable production of h-albumin and cytochrome P450 (CYP) 3A activities for at least 21 days. The mRNA expression levels of 10 of 13 CYP, UDP-glucuronosyltransferase (UGT), and transporters were maintained at >10% of the levels of freshly isolated cFHHs after 21 days. From 1 week, many bile canaliculi were observed between cFHHs, and the accumulation of the multidrug resistance-associated protein and bile salt export pump substrates in these bile canaliculi was clearly inhibited by cyclosporin A. Microarray analysis of cFHHs cultured at high density and at low density (0.53 × 10 5 cells/cm 2 ) revealed that high density culture maintained high expressions of some transcription factors (HNF4α, PXR, and FXR) perhaps involved in the high CYP, UGT and transporter gene expressions of cFHHs. These results strongly suggest that cFHHs could be a novel in vitro tool for drug development studies.