ABSTRACT
<p><b>BACKGROUND</b>To test susceptibility of human liver cell line Hep G2 to HCV in vitro.</p><p><b>METHODS</b>Hep G2 was cultivated with the serum from a chronic hepatitis C patient. After inoculation, plus and minus strand of HCV RNA, the expression of HCV NS3 antigen and the location of HCV RNA in cell and/or supernatant were examined by RT-PCR, immunohistochemistry and in situ hybridization, respectively.</p><p><b>RESULTS</b>HCV RNA could be detected from day 2 to day 40 post-inoculation in both cell and supernatant. HCV NS3 antigen could be expressed in infected cells and HCV RNA was mainly situated within cytoplasm.</p><p><b>CONCLUSIONS</b>The results suggested that HepG2 cell line was not only susceptible to HCV but also could support its replication in vitro.</p>
Subject(s)
Humans , Carcinoma, Hepatocellular , Pathology , Virology , Coculture Techniques , Hepacivirus , Genetics , Physiology , Hepatitis C, Chronic , Virology , Liver Neoplasms , Pathology , Virology , RNA, Viral , Genetics , Tumor Cells, Cultured , Viral Nonstructural Proteins , Genetics , Virus ReplicationABSTRACT
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 NS3 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-NS3 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.
ABSTRACT
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 NS3 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-NS3 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.