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

Autor: Chise Tateno, Chihiro Yamasaki, Yumiko Iwasaki, Yasumi Yoshizane, Ami Yanagi, Yuji Ishida, Yutaka Kageyama, Seiichi Ishida, Kazuaki Chayama, Yuko Ogawa, Yuha Kojima
Rok vydání: 2020
Předmět:
0301 basic medicine
Male
Physiology
Microarrays
Enzyme Metabolism
Gene Expression
Mice
SCID
030226 pharmacology & pharmacy
Biochemistry
Mice
0302 clinical medicine
Drug Metabolism
Animal Cells
Gene expression
Medicine and Health Sciences
Bile
Cytochrome P-450 CYP3A
Glucuronosyltransferase
Enzyme Chemistry
Child
Cells
Cultured
Pregnane X receptor
Multidisciplinary
biology
Chemistry
Body Fluids
Bioassays and Physiological Analysis
Liver
Child
Preschool
Cyclosporine
Medicine
Female
Cellular Types
Anatomy
Research Article
Science
Primary Cell Culture
Surgical and Invasive Medical Procedures
Research and Analysis Methods
digestive system
03 medical and health sciences
Digestive System Procedures
Drug Development
Genetics
Animals
Humans
Pharmacokinetics
Secretion
Pharmacology
Transplantation
Transplantation Chimera
Microarray analysis techniques
Bile Canaliculi
Cytochrome P450
Biology and Life Sciences
Transporter
Cell Biology
Organ Transplantation
Bile Salt Export Pump
Molecular biology
In vitro
Liver Transplantation
030104 developmental biology
biology.protein
Enzymology
Hepatocytes
Physiological Processes
Drug metabolism
Transcription Factors
Zdroj: PLoS ONE
PLoS ONE, Vol 15, Iss 9, p e0237809 (2020)
ISSN: 1932-6203
Popis: 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.
Databáze: OpenAIRE
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