The study reports the establishment of a robust cell culture model for hepatitis C virus (HCV) infection, based on the HCV JFH-1 molecular clone and Huh-7-derived cell lines. This system allows the production and propagation of infectious HCV in vitro, overcoming previous limitations such as low infection efficiency and the inability to propagate virus in naïve cells. The system enables the analysis of host-virus interactions, including viral entry, spread, and antiviral responses, which should facilitate the discovery of antiviral drugs and vaccines for HCV. Key findings include the demonstration that Huh-7.5.1 cells, derived from Huh-7 cells, are highly permissive for HCV replication and can produce infectious particles. The virus generated can be serially passaged without loss in infectivity, and its kinetics of infection are similar to those observed in Huh-7.5.1 cells. Additionally, the study shows that the viral envelope protein E2 and the cellular receptor CD81 are crucial for HCV infection, as their antibodies significantly reduce viral infectivity. The robustness of this cell culture model provides a valuable tool for further research on HCV biology and the development of antiviral strategies.The study reports the establishment of a robust cell culture model for hepatitis C virus (HCV) infection, based on the HCV JFH-1 molecular clone and Huh-7-derived cell lines. This system allows the production and propagation of infectious HCV in vitro, overcoming previous limitations such as low infection efficiency and the inability to propagate virus in naïve cells. The system enables the analysis of host-virus interactions, including viral entry, spread, and antiviral responses, which should facilitate the discovery of antiviral drugs and vaccines for HCV. Key findings include the demonstration that Huh-7.5.1 cells, derived from Huh-7 cells, are highly permissive for HCV replication and can produce infectious particles. The virus generated can be serially passaged without loss in infectivity, and its kinetics of infection are similar to those observed in Huh-7.5.1 cells. Additionally, the study shows that the viral envelope protein E2 and the cellular receptor CD81 are crucial for HCV infection, as their antibodies significantly reduce viral infectivity. The robustness of this cell culture model provides a valuable tool for further research on HCV biology and the development of antiviral strategies.