title: Primary cultures of human hepatocytes isolated from hepatitis C virus-infected cirrhotic livers as a model to study hepatitis C infection.
abstract: OBJECTIVE
  Since the discovery of hepatitis C virus (HCV), researchers have encountered difficulties with in vitro models. The aim of this study was to determine whether HCV-infected human primary hepatocytes, isolated from cirrhotic livers at liver transplantation, can be used as a model to study HCV infection.
  
  
  METHODS
  Hepatocytes were isolated with collagenase and cultured over a 20-day period on different matrices. Viral kinetics was monitored with/without treatment by real-time polymerase chain reaction.
  
  
  RESULTS
  Cell yield and viability were higher with uninfected/non-cirrhotic livers (77.2+/-1.8%) in comparison with HCV-infected cirrhotic livers (68.8+/-12%). HCV-infected hepatocytes behaved similar to non-infected cells and expressed albumin and cytochrome P4502E1. HCV-positive strand was identified in supernatants and cell lysates. HCV-negative strand was only found inside cells and correlated with viral RNA recovery in the medium. Improvement in the degree of hepatocyte differentiation was associated with better HCV recovery. Antiviral treatment with interferon-alpha, EX4 and cyclosporine A induced significant reductions in HCV RNA.
  
  
  CONCLUSIONS
  Primary cultures of HCV-infected human hepatocytes from end-stage cirrhotic livers is feasible, represents an excellent model to study specific virus-host interactions and can be used to assess viral replication.
authors.affiliation: Laboratoire d'hépatologie cellulaire, Centre hospitalier de l'Université de Montréal (CHUM)-Hôpital Saint-Luc, Montréal, QC, Canada.
authors.forename: Valérie-Ann
authors.initials: VA
authors.lastname: Raymond
authors.forename: Subajini
authors.initials: S
authors.lastname: Selliah
authors.forename: Chantal
authors.initials: C
authors.lastname: Ethier
authors.forename: Robert
authors.initials: R
authors.lastname: Houle
authors.forename: Loubna
authors.initials: L
authors.lastname: Jouan
authors.forename: Thibaut
authors.initials: T
authors.lastname: Maniere
authors.forename: Daniel
authors.initials: D
authors.lastname: Lamarre
authors.forename: Bernard
authors.initials: B
authors.lastname: Willems
authors.forename: Marc
authors.initials: M
authors.lastname: Bilodeau
mesh_terms: D000704:Analysis of Variance; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics* / Q000502:physiology; D006526:Hepatitis C / Q000821:virology*; D022781:Hepatocytes / Q000187:drug effects / Q000821:virology*; D054884:Host-Pathogen Interactions; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D008099:Liver / Q000166:cytology* / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication / Q000187:drug effects
chemical_list: D017931:DNA Primers; D016898:Interferon-alpha; D016572:Cyclosporine