Matrigel-embedded 3D culture of Huh-7 cells as a hepatocyte-like polarized system to study hepatitis C virus cycle

Virology. 2012 Mar 30;425(1):31-9. doi: 10.1016/j.virol.2011.12.021. Epub 2012 Jan 24.


Hepatocytes are highly polarized cells where intercellular junctions, including tight junctions (TJs), determine the polarity. Recently, the TJ-associated proteins claudin-1 and occludin have been implicated in hepatitis C virus (HCV) entry and spread. Nevertheless, cell line-based experimental systems that exhibit hepatocyte-like polarity and permit robust infection and virion production are not currently available. Thus, we sought to determine whether cell line-based, Matrigel-embedded cultures could be used to study hepatitis C virus (HCV) infection and virion production in a context of hepatocyte-like polarized cells. In contrast to standard bidimensional cultures, Matrigel-cultured Huh-7 cells adopted hepatocyte polarization features forming a continuous network of functional proto-bile canaliculi structures. These 3D cultures supported HCV infection by JFH-1 virus and produced infective viral particles which shifted towards lower densities with higher associated specific infectivity. In conclusion, our findings describe a novel use of Matrigel to study the entire HCV cycle in a more relevant context.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bile Canaliculi / ultrastructure
  • Biocompatible Materials / chemistry*
  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Cell Polarity
  • Collagen / chemistry*
  • Drug Combinations
  • Hepacivirus / genetics
  • Hepacivirus / metabolism
  • Hepacivirus / pathogenicity*
  • Hepacivirus / ultrastructure
  • Hepatitis C / virology
  • Hepatocytes / metabolism
  • Hepatocytes / virology
  • Humans
  • Laminin / chemistry*
  • Microscopy, Confocal
  • Proteoglycans / chemistry*
  • Tight Junctions / ultrastructure
  • Tissue Embedding / methods*
  • Virion / metabolism*
  • Virion / ultrastructure


  • Biocompatible Materials
  • Drug Combinations
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen