Objective：To establish a sodium taurocholate cotransporting polypeptide（NTCP）-complemented Huh7.5.1 cell line（Huh7.5.1-hNTCP）that supports hepatitis B virus（HBV） and hepatitis C virus（HCV） co-infection. Methods：Huh7.5.1 cells were infected with GV358 recombinant lentiviral vector carrying human NTCP encoding gene，green fluorescent protein（GFP）gene and puromycin resistance gene with the multiplicity of infection（MOI）of 50. NTCP positive cells were obtained through the puromycin screening and NTCP expression was identified by Western blotting. Huh7.5.1-hNTCP cells were infected with HBV derived from HepAD38 cells. The expression of HBsAg，HBeAg and HBV DNA at different time points were detected by ELISA and qPCR. HCV-JFH1 was added 24 hours after HBV infection，and then HBV DNA and HCV RNA were detected in co-infected cells. Results：Western blotting results indicated that Huh7.5.1-hNTCP cell line stably expressed the HBV receptor NTCP. ELASA and qPCR results showed that HBsAg，HBeAg and HBV DNA secretion increased gradually and peaked on the 9th day in HBV infected Huh7.5.1-hNTCP cells. The results of qPCR suggested that the replication of HBV DNA and HCV RNA increased gradually at different time points in HBV and HCV co-infected cells. Conclusion：Huh7.5.1-hNTCP cell line was established，and it can be co-infected by HBV and HCV.