RT Journal Article SR Electronic T1 Immortalization of human zone I hepatocytes from biliary atresia with CDK4R24C, cyclin D1, and TERT for cytochrome P450 induction testing JF bioRxiv FD Cold Spring Harbor Laboratory SP 729525 DO 10.1101/729525 A1 Nishiwaki, Manami A1 Toyoda, Masashi A1 Oishi, Yoshie A1 Ishida, Seiichi A1 Horiuchi, Shin-ichiro A1 Makino, Hatsune A1 Kimura, Tohru A1 Ohno, Shin-ichi A1 Ohkura, Takashi A1 Enosawa, Shin A1 Akutsu, Hidenori A1 Nakazawa, Atsuko A1 Kasahara, Mureo A1 Kiyono, Tohru A1 Umezawa, Akihiro YR 2019 UL http://biorxiv.org/content/early/2019/08/08/729525.abstract AB Background 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.Methods 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.Results 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.Conclusions 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.