Abstract: Objective To establish a hybrid cell different from either HepG₂ cell or human primary hepatic cell which carries HBV and can be serial subcultivation in vitro. MethodsHGPRT^-HepG₂ cell, which was induced by 6MP, was fused with human primary hepatic cell infected by hepatitis B virus. After being screened with HAT, the hybrid cells were cloned through limiting dilution assay. Subsequently,the hybrid cells were identified by karyotype analysis. HBV DNA and HBsAg、nd HBeAg were determined at different generations. HepG₂ and human primary hepatic cell infected by hepatitis B virus were taken as controls. ResultsHybrid cells were morphologically similar to HGPRT-HepG₂cells and can be subcultured in vitro. The Karyotype analysis showed that the modal chromosome number was 99 in the hybrid cells, indicating that the hybrid cells contained all genomic factors from both HepG₂ and human hepatic cells. HBV DNA can be detected in the cells and supernate by nest PCR. In the supernate, HBsAg and HBeAg can be detected. Conceivably the hybrid cells possessed the replication and generation in vitro of HepG₂ as well as the sensitivity of human primary hepatic cell to HBV, which paved secretion of HBV DNA. Conclusion As a new hybrid cell line, it has the characteristics of both ways for further study of HBV infection cell model.

Key words: HepG₂ cell, HGPRT mutation, Human primary hepatic cell, Cell fusion, Hybrid cell