Primary hepatocyte culture supports hepatitis C virus replication: a model for infection-associated hepatocarcinogenesis

Hepatology. 2010 Jun;51(6):1922-32. doi: 10.1002/hep.23616.


Analysis of progressive changes in hepatic gene expression that underlie hepatocarcinogenesis following hepatitis C virus (HCV) infection require examination of long-term cultures of normally differentiating primary human hepatocytes. We report a culture system of primary hepatocytes that support productive replication of infectious HCV. Hepatic functions were analyzed by reverse-transcription polymerase chain reaction amplification of total cell RNA from cultures maintained in serum-free defined medium for up to 190 days. Sustained hepatic function was assessed by expression of albumin, alpha-fetoprotein, cytochrome P4502E1, cytokeratin-18, type-1 collagen, transforming growth factor-beta 1, matrix metalloproteinase-2 (MMP-2), MMP-13, and interferon alpha-receptors 1 and 2. Normally differentiated human primary hepatocytes supported productive replication of infectious clones of HCV genotypes 1a, 1b, and 2a; virus infection was inhibited by antibodies against CD81 virus entry factor. Virus released into the culture media of HCV-infected primary hepatocytes repeatedly passage to naïve hepatocytes. Replication of the three HCV genotypes shows interferon sensitivity observed in natural infections.

Conclusion: Sustained cultures of physiologic host cells for the propagation of infectious HCV strains should accelerate studies of host response to HCV infection and progressive liver disease.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Line
  • Coculture Techniques
  • Culture Media
  • Genome, Viral
  • Hepacivirus / physiology*
  • Hepatocytes / cytology
  • Hepatocytes / metabolism
  • Hepatocytes / virology*
  • Humans
  • Interferon-alpha
  • Liver Neoplasms / virology
  • RNA, Viral / biosynthesis*
  • Rats
  • Virus Release
  • Virus Replication*


  • Culture Media
  • Interferon-alpha
  • RNA, Viral