This study addresses the lack of a reliable cell culture system for hepatitis C virus (HCV) replication, a major cause of chronic liver disease. The authors successfully generated infectious pseudo-particles by expressing unmodified and functional HCV glycoproteins (E1 and E2) onto retroviral and lentiviral core particles. These pseudo-particles, containing a green fluorescent protein (GFP) marker, allowed for the rapid and reliable determination of infectivity. Primary hepatocytes and hepatocarcinoma cells were found to be the primary targets of infection in vitro. High infectivity required both E1 and E2 glycoproteins, and was neutralized by sera from HCV-infected patients and some anti-E2 monoclonal antibodies. The study also investigated the role of putative HCV receptors, finding that neither LDLr nor CD81 was sufficient for HCV entry. The pseudo-particles mimic the early infection steps of parental HCV and are suitable for developing new antiviral therapies.This study addresses the lack of a reliable cell culture system for hepatitis C virus (HCV) replication, a major cause of chronic liver disease. The authors successfully generated infectious pseudo-particles by expressing unmodified and functional HCV glycoproteins (E1 and E2) onto retroviral and lentiviral core particles. These pseudo-particles, containing a green fluorescent protein (GFP) marker, allowed for the rapid and reliable determination of infectivity. Primary hepatocytes and hepatocarcinoma cells were found to be the primary targets of infection in vitro. High infectivity required both E1 and E2 glycoproteins, and was neutralized by sera from HCV-infected patients and some anti-E2 monoclonal antibodies. The study also investigated the role of putative HCV receptors, finding that neither LDLr nor CD81 was sufficient for HCV entry. The pseudo-particles mimic the early infection steps of parental HCV and are suitable for developing new antiviral therapies.