人胎肝细胞体外感染丙型肝炎病毒的定位研究

Orientation Researches of Human Fetal Hepatocytes Infected with Hepatitis C Virus in vitro

为了证实人胎肝细胞在体外与丙型肝炎病毒 (HCV)阳性血清共培养 2 4h是否能感染 HCV病毒而作定位研究。采用透射电镜、免疫电镜的手段对于 HCV阳性血清共培养的人胎肝细胞中病毒样颗粒进行定位研究 ,以测定细胞内是否有 HCV。结果发现在透视电镜下人胎肝细胞胞浆中有很多病毒样颗粒 ,直径约 45 nm。通过免疫电镜进一步证实在人胎肝细胞中这些病毒样颗粒含有 HCV基因表达产物 NS3及 NS5。此外 ,人胎肝细胞在体外与 HCV阳性血清共培养 2 4h并经多次洗涤后的培养上清加入到新鲜分离的正常人胎肝细胞培养基中继续培养 ,可以得出上述相同的结果。这些结果表明人胎肝细胞在体外培养情况下可以感染 HCV,并且感染了

Orientation researches were carried out to confirm whether human fetal hepatocytes (HFH) might be infected with hepatitis C virus (HCV) after co culturing with HCV positive serum in vitro for 24 h. Under transmission and immune electron microscopy, the virus like particles in the HFH co cultured with HCV positive serum were located to determinate whether there were HCV in the cells. It was found that there were lots of virus like particles with about 45 nm in diameter in cytoplasm of the HFH under transmission electron microscopy. It was further confirmed that these virus like particles in HFH contained HCV gene expression products NS3 and NS5 under immune electron microscopy. Moreover, the culture supernatant of HFH co cultured with HCV positive serum for 24 h in vitro with many times of washing was added into the culture medium of the HFH separated freshly and continued to culture, the same results mentioned above were obtained. These results indicated that HFH may be infected with HCV in the circumstances of culture in vitro and the HFH infected with HCV can produce contagious HCV particles to secrete into the culture supernatant.