HBV携带者肝细胞体外培养的研究

Study on hepatitis B virus replication in human primary hepatic cell with hepatitis B virus in vitro

目的:体外培养HBV携带者人原代肝细胞,研究细胞内HBV的复制情况及持续时间,为临床抗病毒试验治疗奠定基础。方法:分离HBV携带者人原代肝细胞进行体外培养。14d后,测绘细胞的生长曲线和细胞核型分析。分别用巢式PCR和ELISA法对细胞匀浆及培养上清液进行HBV-DNA和HBsAg、HBeAg的检测。结果:HBV携带者人原代肝细胞和培养上清液可检测到HBV-rcDNA,培养上清液内检测到HBsAg和HBeAg。细胞培养3周时开始出现大量死亡。结论:该细胞体外培养期间仍能保持对HBV感染和复制的特性,为体外研究HBV感染情况奠定了基础。

Objective To research the survivin of hepatitis B virus carried in human primary hepatocytes in vitro and the lasting of the cells. Methods The human primary hepatic cells carrying hepatitis B virus were isolated in vitro. After 14 days, supernates and cells of culture were examined for HBV-DNA, HBsAg and HBeAg by PCR and ELISA respectively. Results HBV reDNA was detected in supernates and cells. HBsAg and HBeAg were detected in supernates. After 3 weeks, a great smount of cells began dying. Conclusion The human primary hepatic cells carrying hepatitis B virus keep the characters of infection and replication of hepatitis B virus during culture in vitro. The findings may make foundations for studying hepatitis B virus infection in vitro.