摘要
目的 研究丙型肝炎病毒 (HCV)在体外培养原代人胎肝细胞和人肝癌细胞系 772 1中复制和表达的异同 ,探讨HCV体外培养条件。方法 原代人胎肝细胞和人肝癌细胞系 772 1分别与HCV感染血清共孵育后 ,用逆转录 聚合酶链反应、原位杂交、免疫组化检测细胞和培养上清中的HCVRNA和抗原表达。结果 从孵育的第 2～ 3天 ,即可在细胞内和 /或培养上清中间断地检出HCVRNA(其中HCV在 772 1细胞株中的复制至少可达 66d ,HCV在人胎肝细胞中的复制持续 2 5d) ;HCV抗原可在感染细胞内得到稳定表达 ,感染细胞内存在HCV负链RNA杂交信号 ,且多位于细胞浆。结论 两种肝细胞对HCV易感 ,尤其是 772 1细胞可以稳定地支持HCV体外复制 ,可用于HCV体外长期培养的靶细胞。
Objective To study the similarities and differences on in vitro replication and expression of hepatitis C virus (HCV) between human fetal hepatocytes (HFH) and 7721 cell line. Methods Human fetal hepatocytes and a hepatoma cell line 7721 were incubated with a serum from hepatitis C patient. After incubation, the presence of HCV RNA, the expression of HCV NS 3 antigens in cells and/or supernatant were examined by RT PCR, in situ hybridization and immunohistochemistry, respectively. Results It was found that: ①The intracellular HCV RNA was first detected on d 2～3 post incubation and then could be intermittently detected in cells and/or supernatant subsequently (HCV RNA could be detected in 7721 cells during a period of at least 66 days. In HFH, HCV RNA could be detected up to 25 days after incubation); ②HCV NS 3 antigen could be expressed in infected cells; ③Minus strand RNA of HCV was mainly located within cytoplasm by in situ hybridization. Conclusion The results suggest that both the fetal hpatocytes and the hepatoma cell line 7721 are susceptible to HCV, and especially 7721 cell line can stably support HCV replication in vitro and may be used as the target cell for long term cultures of HCV.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2001年第2期204-207,共4页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目!( 396 70 6 72 )
