title: Establishment and characterization of immortalized human hepatocyte cell line for applications in bioartificial livers.
abstract: An immortalized human hepatocyte cell line, HepLi5, was established via transfection of Simian virus 40 large T antigen (SV40 LT) into primary human hepatocytes. The morphologic and functional characteristics of HepLi5 cells were evaluated. The expression of SV40 LT in HepLi5 cells was detected by reverse transcription-PCR (RT-PCR) and western blotting. mRNA expression of liver-enriched genes, including glutamine synthetase, albumin, and cytochrome P450 was detected via RT-PCR in HepLi5 cells. Activity of CYP1A2, one of the drug-metabolizing P450 enzymes, was detected. Subcutaneous injection of HepLi5 cells into nude mice did not induce tumors within 3 months. Short Tandem Repeat results confirmed the authenticity of the cell line. Clinical-grade quantities of HepLi5 cells could be harvested using large-scale culture in roller bottles after which their cellular function was significantly enhanced. Therefore, the immortalized HepLi5 cells are a suitable cell source for applications in bioartificial livers.
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China. xiaopingpan@zju.edu.cn
authors.forename: XiaoPing
authors.initials: X
authors.lastname: Pan
authors.forename: JianZhou
authors.initials: J
authors.lastname: Li
authors.forename: WeiBo
authors.initials: W
authors.lastname: Du
authors.forename: XiaoPeng
authors.initials: X
authors.lastname: Yu
authors.forename: ChunXia
authors.initials: C
authors.lastname: Zhu
authors.forename: ChengBo
authors.initials: C
authors.lastname: Yu
authors.forename: HongCui
authors.initials: H
authors.lastname: Cao
authors.forename: YiMin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.forename: LanJuan
authors.initials: L
authors.lastname: Li
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002472:Cell Transformation, Viral; D022781:Hepatocytes / Q000502:physiology*; D006801:Humans; D019164:Liver, Artificial; D051379:Mice; D008819:Mice, Nude; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral