Komorowski Marcin, Kołakowska Agnieszka, Godzik Paulina, Madaliński Kazimierz
Pracownia Immunopatologii Zakazen Hepatotropowych, Zakładu Wirusologii Narodowego Instytutu Zdrowia Publicznego--Państwowego Zakładu Higieny w Warszawie.
Med Dosw Mikrobiol. 2012;64(3):239-44.
Infection with hepatitis C virus is a serious worldwide health problem. Since its discovery in 1989, the development of a cell culture system for HCV has been a major goal for scientists worldwide. In 2005 the first tissue culture that led to the production of HCV particles (2a genotype) has been created. The aim of the study was to determine viral RNA level in an infectious HCV cell culture system.
Huh-7.5 cell line was infected with the JFH1 (Japanese Fulminant Hepatitis) RNA by lipofection. The level of HCV RNA was measured in supernatant of the cell culture by Real-Time PCR method.
HCV RNA was detected in the supernatant of Huh-7.5 cell line infected with the use ofJFH 1 RNA and lipofectamin. The maximal level of HCV RNA (3.69 x 10(9) copies/ml) was detected 96h after transfection. After about 30 days oftransfection, HCV RNA was on the stable level (10(7)-10(8)).
Huh-7.5 cell line infected with HCV form a robust cell culture model of HCV infection. HCV replicates on high levels in Huh-7.5, which is the crucial event for the investigation of new antivirals in in vitro model.
丙型肝炎病毒感染是一个严重的全球性健康问题。自1989年被发现以来，建立丙型肝炎病毒细胞培养系统一直是全球科学家的主要目标。2005年，首个能产生丙型肝炎病毒颗粒（2a基因型）的组织培养物被创建出来。本研究的目的是测定感染性丙型肝炎病毒细胞培养系统中的病毒RNA水平。
采用脂质体转染法将JFH1（日本暴发性肝炎）RNA感染Huh-7.5细胞系。通过实时聚合酶链反应法测定细胞培养上清液中的丙型肝炎病毒RNA水平。
在使用JFH1 RNA和脂质体转染试剂感染的Huh-7.5细胞系的上清液中检测到丙型肝炎病毒RNA。转染后96小时检测到丙型肝炎病毒RNA的最高水平（3.69×10⁹拷贝/毫升）。转染约30天后，丙型肝炎病毒RNA处于稳定水平（10⁷ - 10⁸）。
感染丙型肝炎病毒的Huh-7.5细胞系形成了一个强大的丙型肝炎病毒感染细胞培养模型。丙型肝炎病毒在Huh-7.5细胞中大量复制，这是在体外模型中研究新型抗病毒药物的关键事件。
