Institution: | a Molecular Biology Unit, Hospital Universitario de la Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid 28006, Spainb CIBERehd, Instituto de Salud Carlos III, Madrid 28029, Spainc Université de Lyon, UCB-Lyon1, IFR128, Lyon F-69007, Franced INSERM, U758, Lyon F-69007, Francee Ecole Normale Supérieure de Lyon, Lyon F-69007, Francef Laboratory of Experimental Hepatology and Drug Targeting, Universidad de Salamanca, Salamanca 37007, Spaing Research Unit, Hospital Universitario de Salamanca, Salamanca 37007, Spainh Liver Unit, Hospital Universitario de la Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid 28006, Spaini Division of Gene Therapy and Hepatology, Centro de Investigación en Medicina Aplicada, CIMA, Pamplona 31008, Spainj Inserm U748, Université de Strasbourg, F-67000 Strasbourg, Francek Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Madrid 28049, Spain |
Abstract: | 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. |