title: Production of infectious hepatitis C virus in primary cultures of human adult hepatocytes.
abstract: OBJECTIVE
  Although hepatitis C virus (HCV) can be grown in the hepatocarcinoma-derived cell line Huh-7, a cell-culture model is needed that supports its complete, productive infection cycle in normal, quiescent, highly differentiated human hepatocytes. We sought to develop such a system.
  
  
  METHODS
  Primary cultures of human adult hepatocytes were inoculated with HCV derived from Huh-7 cell culture (HCVcc) and monitored for expression of hepatocyte differentiation markers and replication of HCV. Culture supernatants were assayed for HCV RNA, core antigen, and infectivity titer. The buoyant densities of input and progeny virus were compared in iodixanol gradients.
  
  
  RESULTS
  While retaining expression of differentiation markers, primary hepatocytes supported the complete infectious cycle of HCV, including production of significant titers of new infectious progeny virus, which was called primary-culture-derived virus (HCVpc). Compared with HCVcc, HCVpc had lower average buoyant density and higher specific infectivity; this was similar to the characteristics of virus particles associated with the very-low-density lipoproteins that are produced during in vivo infection. These properties were lost after re-culture of HCVpc in poorly differentiated Huh-7 cells, suggesting that authentic virions can be produced only by normal hepatocytes that secrete authentic very-low-density lipoproteins.
  
  
  CONCLUSIONS
  We have established a cell-culture-based system that allows production of infectious HCV in physiologically relevant human hepatocytes. This provides a useful tool for the study of HCV interactions with its natural host cell and for the development of antiviral therapies.
authors.affiliation: INSERM, Equipe Avenir Virologie de l'Hépatite C, Institut Cochin, Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Podevin
authors.forename: Arnaud
authors.initials: A
authors.lastname: Carpentier
authors.forename: Véronique
authors.initials: V
authors.lastname: Pène
authors.forename: Lynda
authors.initials: L
authors.lastname: Aoudjehane
authors.forename: Matthieu
authors.initials: M
authors.lastname: Carrière
authors.forename: Sakina
authors.initials: S
authors.lastname: Zaïdi
authors.forename: Céline
authors.initials: C
authors.lastname: Hernandez
authors.forename: Vanessa
authors.initials: V
authors.lastname: Calle
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Méritet
authors.forename: Olivier
authors.initials: O
authors.lastname: Scatton
authors.forename: Marlène
authors.initials: M
authors.lastname: Dreux
authors.forename: François-Loïc
authors.initials: FL
authors.lastname: Cosset
authors.forename: Takaji
authors.initials: T
authors.lastname: Wakita
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
authors.forename: Sylvie
authors.initials: S
authors.lastname: Demignot
authors.forename: Filoména
authors.initials: F
authors.lastname: Conti
authors.forename: Arielle R
authors.initials: AR
authors.lastname: Rosenberg
authors.forename: Yvon
authors.initials: Y
authors.lastname: Calmus
mesh_terms: D000328:Adult; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D016679:Genome, Viral; D016174:Hepacivirus / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D014779:Virus Replication