丙型肝炎病毒体外感染原代人胎肝细胞的研究

目的　研究丙型肝炎病毒 (HCV)体外细胞培养方法。方法　原代人胎肝细胞与HCV感染血清共孵育后 ,用逆转录 聚合酶链反应、原位杂交、免疫组化分别检测细胞和培养上清中的HCVRNA和HCV抗原表达。结果　从感染血清和细胞共同孵育后的 2～ 2 5d ,细胞内和 /或培养上清中可间断检出HCV正、负链RNA ;HCVNS3抗原在细胞内能稳定表达 ;原位杂交显示HCV负链RNA阳性物质多位于细胞浆。结论　原代人胎肝细胞对HCV易感 ,可用作HCV体外感染和复制的靶细胞

Study on in vitro infection of human fetal hepatocytes by hepatitis C virus

Objective Cultivation of hepatitis C virus in vitro . Methods Human fetal hepatocytes were incubated with serum from a hepatitis C patient. After incubation, the presence of HCV RNA, the expression of HCV NS3 antigens in cells and/or supernatant were detected by RT PCR, in situ hybridization and immunohistochemistry respectively. Results It was found that plus and minus strand of HCV RNA could be detected in cells and/or supernatant during a period of 25 days post inoculation. HCV NS3 antigens were expressed within cytoplasm of infected cells. HCV RNA was located within cytoplasm. Conclusion Human fetal hepatocytes were susceptible to HCV and could be used as target cell for the infection and replication of HCV in vitro.