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Tytuł pozycji:

Culture 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.

Tytuł :
Culture 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.
Autorzy :
Yamasaki C; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Ishida Y; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.; Research Center for Hepatology and Gastroenterology, Hiroshima University, Hiroshima, Japan.
Yanagi A; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Yoshizane Y; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Kojima Y; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Ogawa Y; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Kageyama Y; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Iwasaki Y; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.
Ishida S; Department of Pharmacology, National Institute of Health Sciences, Kanagawa, Japan.
Chayama K; Department of Gastroenterology and Metabolism, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Higashihiroshima, Hiroshima, Japan.
Tateno C; PhoenixBio Co., Ltd., Higashi-Hiroshima, Hiroshima, Japan.; Research Center for Hepatology and Gastroenterology, Hiroshima University, Hiroshima, Japan.
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Źródło :
PloS one [PLoS One] 2020 Sep 11; Vol. 15 (9), pp. e0237809. Date of Electronic Publication: 2020 Sep 11 (Print Publication: 2020).
Typ publikacji :
Journal Article; Research Support, Non-U.S. Gov't
Język :
English
Imprint Name(s) :
Original Publication: San Francisco, CA : Public Library of Science
MeSH Terms :
Transplantation Chimera*
Bile Canaliculi/*drug effects
Drug Development/*methods
Hepatocytes/*drug effects
Primary Cell Culture/*methods
Animals ; Bile Canaliculi/cytology ; Bile Canaliculi/metabolism ; Cells, Cultured ; Child ; Child, Preschool ; Cyclosporine/pharmacology ; Cytochrome P-450 CYP3A/genetics ; Cytochrome P-450 CYP3A/metabolism ; Female ; Glucuronosyltransferase/genetics ; Glucuronosyltransferase/metabolism ; Hepatocytes/cytology ; Hepatocytes/metabolism ; Humans ; Male ; Mice ; Mice, SCID ; Transcription Factors/genetics ; Transcription Factors/metabolism
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Substance Nomenclature :
0 (Transcription Factors)
83HN0GTJ6D (Cyclosporine)
EC 1.14.14.1 (Cytochrome P-450 CYP3A)
EC 2.4.1.17 (Glucuronosyltransferase)
Entry Date(s) :
Date Created: 20200911 Date Completed: 20201021 Latest Revision: 20201021
Update Code :
20210210
PubMed Central ID :
PMC7485858
DOI :
10.1371/journal.pone.0237809
PMID :
32915792
Czasopismo naukowe
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 × 105 cells/cm2) 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 × 105 cells/cm2) 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.
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