Matrigel-embedded 3D culture of Huh-7 cells as a hepatocyte-like polarized system to study hepatitis C virus cycle
Matrigel-embedded 3D culture of Huh-7 cells as a hepatocyte-like polarized system to study hepatitis C virus cycle
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DOI：
10.1016/j.virol.2011.12.021
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发表时间：
2012-03-30
期刊：
影响因子：
3.7
通讯作者：
Majano, Pedro L.
中科院分区：
文献类型：
--
作者：
Molina-Jimenez, Francisca;Benedicto, Ignacio;Majano, Pedro L.
肝细胞是高度极化的细胞，其中细胞间连接（包括紧密连接（TJ））决定极性。最近，TJ相关蛋白claudin-1和occludin已被牵连在丙型肝炎病毒（HCV）的进入和传播。然而，目前还没有基于细胞系的实验系统，其表现出肝细胞样极性并允许稳健的感染和病毒体产生。因此，我们试图确定基于细胞系的基质胶包埋培养物是否可用于研究丙型肝炎病毒（HCV）感染和肝细胞样极化细胞背景下的病毒体产生。与标准的二维培养相比，基质胶培养的Huh-7细胞采用肝细胞极化特征，形成功能性原胆小管结构的连续网络。这些3D培养物支持JFH-1病毒的HCV感染，并产生感染性病毒颗粒，其向较低密度转移，具有较高的相关特异性感染性。总之，我们的研究结果描述了一种新的使用基质胶研究整个HCV周期在一个更相关的背景下。皇冠版权所有（C）2012由Elsevier Inc.发布。All rights reserved.
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. Crown Copyright (C) 2012 Published by Elsevier Inc. All rights reserved.