title: Reversible immortalization of human hepatocytes mediated by retroviral transfer and site-specific recombination.
abstract: OBJECTIVE
  To establish a method for the reversible immortalization of human hepatocytes, which may offer a good and safe source of hepatocytes for practical applications.
  
  
  METHODS
  We successfully isolated primary human hepatocytes from surgically resected liver tissue taken from a patient with liver hemangiomas. The freshly isolated cells were then immortalized with retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40T) and hygromycin-resistance genes flanked by paired loxP recombination targets.
  
  
  RESULTS
  The freshly isolated hepatocytes with high viability (85%) were successfully immortalized using retroviral gene transfer of SV40T. SV40T in the immortalized cells was then excised by Cre/loxP site-specific recombination. This cell population exhibited the characteristics of differentiated hepatocytes.
  
  
  CONCLUSIONS
  We successfully established reversibly immortalized human hepatocytes, which will provide an unlimited supply of cells for practical applications.
authors.affiliation: Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Fan-Ying
authors.initials: FY
authors.lastname: Meng
authors.affiliation: Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Feng-Hui
authors.initials: FH
authors.lastname: Yang
authors.affiliation: Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Chun-You
authors.initials: CY
authors.lastname: Li
authors.affiliation: Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.affiliation: Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002472:Cell Transformation, Viral; D002934:Cinnamates / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics; D005822:Genetic Vectors; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology*; D006801:Humans; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics* / Q000378:metabolism; D012709:Serum Albumin / Q000235:genetics / Q000378:metabolism; D000075462:Serum Albumin, Human; D013997:Time Factors; D014161:Transduction, Genetic
chemical_list: C422455:ALB protein, human; D000900:Anti-Bacterial Agents; D000952:Antigens, Polyomavirus Transforming; D002934:Cinnamates; D012709:Serum Albumin; D006921:Hygromycin B; C026273:hygromycin A; C045073:Cre recombinase; D019426:Integrases; D000075462:Serum Albumin, Human
keywords: Hepatocyte; Hepatocyte isolation; Primary human hepatocytes; Reversible immortalization; SV40T