IN-VITRO INFECTION OF ADULT NORMAL HUMAN HEPATOCYTES IN PRIMARY CULTURE BY HEPATITIS-C VIRUS

In vitro infection of adult normal human hepatocytes in primary cultur e has been performed for investigating the replication cycle of hepati tis C virus (HCV) in differentiated cells. Hepatocytes were prepared f rom liver tissue resected from donors who tested negative for HCV, and inoculation was performed 3 days after plating with 33 HCV serum samp les of different virus load and genotype, The presence of intracellula r HCV RNA, detected by a strand-specific rTth RT-PCR assay, was used a s evidence of infection. A kinetics analysis of HCV replication reveal ed that intracellular negative-strand RNA appeared at day 1 post-infec tion with a maximum level at days 3 and 5, followed by a decrease unti l day 14. At day 5, we estimated that the copy level of viral RNA was amplified at least 15-fold in infected cells. The level of intracellul ar HCV RNA in response to different serum samples was reproducible fro m one hepatocyte culture to another, suggesting that there is no inter -individual variability in the susceptibility of hepatocytes to HCV in fection. These findings indicate that adult human hepatocytes in prima ry culture retain their susceptibility to in vitro HCV infection and s upport HCV RNA replication. This model should represent a valuable too l for the study of initial steps of the HCV replication cycle and for the evaluation of antiviral molecules.