| Objective(s): According to the World Health Organization(WHO),as of June24,2022,there are approximately 58 million people infected with chronic hepatitis C virus(HCV)worldwide,and there is still no effective vaccine available.Because the pathogenesis and disease course of Rodent Hepacivirus(RHV)infection in rats found in Norway are similar to those of HCV,RHV is currently the most valuable alternative model for vaccine-induced adaptive immunity assessment of hepatitis C.Although RHV animal models and infection-enhanced mutant cell models have been established successively,wild full-length RHV cell infection models are still not available.The aim of this study was to establish a cell line that could be infected by wild full-length RHV to lay the foundation for more accurate vaccine efficacy evaluation.Methods: Rat primary hepatocytes(PRH)were obtained by isolation using the Seglen two-step method,and the infectivity of RHV to PRH was verified by de-infecting PRH with RHV,and further immortalizing PRH with lentivirus SV40 large T to establish the immortalized hepatocyte line RH20 T,which was characterized and verified.Finally,RHV was used to infect RH20 T,and the infectivity of RHV on RH20 T was verified again.The RH20 T was used to establish the RHV titer detection system.Results:(1)The isolated PRH can be infected by RHV.(2)RH20T,which has the ability to store glycogen and lipids as well as synthesize and secrete Alanine Transaminase(ALT),Aspartate Transaminase(AST)and Albumin(ALb),was not contaminated by exogenous cell lines.(3)RH20T was still able to be infected by RHV,and the RHV titer detection system was successfully established.Conclusion(s): The rat hepatocyte cell line susceptible to RHV was successfully established,and this cell model not only provided accurate quantification of virus titers,but also laid the foundation for more accurate evaluation of vaccine effects,which can be used for more in-depth studies in the field of hepatitis virus. |
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