論文

国際誌
2020年10月15日

Immortalization of human hepatocytes from biliary atresia with CDK4R24C, cyclin D1, and TERT for cytochrome P450 induction testing.

Scientific reports
  • Manami Nishiwaki
  • ,
  • Masashi Toyoda
  • ,
  • Yoshie Oishi
  • ,
  • Seiichi Ishida
  • ,
  • Shin-Ichiro Horiuchi
  • ,
  • Hatsune Makino-Itou
  • ,
  • Tohru Kimura
  • ,
  • Shin-Ichi Ohno
  • ,
  • Takashi Ohkura
  • ,
  • Shin Enosawa
  • ,
  • Hidenori Akutsu
  • ,
  • Atsuko Nakazawa
  • ,
  • Mureo Kasahara
  • ,
  • Tohru Kiyono
  • ,
  • Akihiro Umezawa

10
1
開始ページ
17503
終了ページ
17503
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1038/s41598-020-73992-3

Hepatocytes are an important tool for in vitro toxicology testing. In addition to primary cultures, a limited number of immortalized cell lines have been developed. We here describe a new cell line, designated as HepaMN, which has been established from a liver associated with biliary atresia. Hepatocytes were isolated from a liver of 4-year-old girl with biliary atresia and immortalized by inoculation with CSII-CMV-TERT, CSII-CMV-Tet-Off, CSII-TRE-Tight-cyclin D1 and CSII-TRE-Tight-CDK4R24C (mutant CDK4: an INK4a-resistant form of CDK4) lentiviruses at the multiplicity of infection of 3 to 10. HepaMN cells exhibited morphological homogeneity, displaying hepatocyte-like phenotypes. Phenotypic studies in vivo and in vitro revealed that HepaMN cells showed polarized and functional hepatocyte features along with a canalicular cell phenotype under defined conditions, and constitutively expressed albumin and carbamoyl phosphate synthetase I in addition to epithelial markers. Since HepaMN cells are immortal and subcloned, kinetics and expression profiles were independent of population doublings. HepaMN cells showed increased CYP3A4 expression after exposure to rifampicin, implying that their close resemblance to normal human hepatocytes makes them suitable for research applications including drug metabolism studies.

リンク情報
DOI
https://doi.org/10.1038/s41598-020-73992-3
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/33060611
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7567112

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