title: Matrigel-embedded 3D culture of Huh-7 cells as a hepatocyte-like polarized system to study hepatitis C virus cycle.
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.
authors.affiliation: Molecular Biology Unit, Hospital Universitario de la Princesa, Instituto de Investigación Sanitaria Princesa (IP), Madrid 28006, Spain. fmolina.hlpr@salud.madrid.org
authors.forename: Francisca
authors.initials: F
authors.lastname: Molina-Jimenez
authors.forename: Ignacio
authors.initials: I
authors.lastname: Benedicto
authors.forename: Viet Loan
authors.initials: VL
authors.lastname: Dao Thi
authors.forename: Virginia
authors.initials: V
authors.lastname: Gondar
authors.forename: Dimitri
authors.initials: D
authors.lastname: Lavillette
authors.forename: Jose J
authors.initials: JJ
authors.lastname: Marin
authors.forename: Oscar
authors.initials: O
authors.lastname: Briz
authors.forename: Ricardo
authors.initials: R
authors.lastname: Moreno-Otero
authors.forename: Rafael
authors.initials: R
authors.lastname: Aldabe
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Baumert
authors.forename: François-Loïc
authors.initials: FL
authors.lastname: Cosset
authors.forename: Manuel
authors.initials: M
authors.lastname: Lopez-Cabrera
authors.forename: Pedro L
authors.initials: PL
authors.lastname: Majano
mesh_terms: D001648:Bile Canaliculi / Q000648:ultrastructure; D001672:Biocompatible Materials / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D016764:Cell Polarity; D003094:Collagen / Q000737:chemistry*; D004338:Drug Combinations; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D006526:Hepatitis C / Q000821:virology; D022781:Hepatocytes / Q000378:metabolism / Q000821:virology; D006801:Humans; D007797:Laminin / Q000737:chemistry*; D018613:Microscopy, Confocal; D011509:Proteoglycans / Q000737:chemistry*; D019108:Tight Junctions / Q000648:ultrastructure; D016610:Tissue Embedding / Q000379:methods*; D014771:Virion / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D001672:Biocompatible Materials; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen