人胎肝细胞体外感染HCV的定性研究

Qualitative Research of Human Fetal Hepatocytes Infected with Hepatitis C Virus in Vitro

目的:通过定性研究探讨人胎肝细胞在体外与丙型肝炎病毒(HCV)阳性血清共培养24h是否能感染HCV。方法:人胎肝细胞在体外与HCV阳性血清共培养24h后,采用原位RT-PCR技术对其进行HCV定性检测。结果:通过原位RT-PCR技术检测,发现人胎肝细胞内HCV正、负链RNA均为阳性。此外,将人胎肝细胞在体外与HCV阳性血清共培养24h并经多次洗涤后继续培养的上清液加入到新鲜分离的正常人胎肝细胞培养基中培养,可以得到上述相同的鲒果。结论:人胎肝细胞在体外培养情况下可以感染HCV,并支持HCV在其内复制,产生具有传染性的HCV颗粒分泌到培养液中。

Aim: Qualitative researches were carried out to confirm whether human fetal hepatocytes (HFH) were infected with hepatitis c viruses (HCV) after co-culturing with HCV positive serum in vitro for 24 hours. Methods: The HFH co-cultured with HCV positive serum were carried out the qualitative researches by means of in situ RT-PCR technique to determinate whether there were HCV RNA in the cells. Results: It was found that the plus and minus strands of HCV RNA in the cells were all positive, after examinated by in situ RT-PCR technique. Moreover, the culture supernatant of HFH co-culturing with HCV positive serum for 24 hours and through washing for many times was added into the culture medium of HFH separated freshly and continued to culture, the same results mentioned above might be arrived at. Conclusions; HFH may be infected with HCV in the circumstances of culture in vitro aid support HCV replicating in the cells to produce contagious HCV particles to secrete into the culture supernatant arc und HFH.