Culture and characterization of human hepatocytes isolated from hepatitis C virus infected liver: Effect of collagen-coated surface and FBS-supplemented medium

The aim of this study is to maintain and prolong the viability and metabolic functions of hepatocytes isolated from hepatitis C virus (HCV)-infected human livers. Collagen-coated surface and 10% fetal bovine serum (FBS) supplemented culture media were studied. Metabolic functions and attachment ability of human hepatocytes in these conditions were compared with normalized conditions (Petri dish, medium without FBS). Our results showed that collagen-coated dishes with 10% FBS-supplemented culture medium increased urea production and albumin synthesis. Urea concentration remained stable (30 pg/cell/h) during the first 3-days when human hepatocytes were cultured in collagen-coated dishes. These hepatocytes showed higher urea production when compared with hepatocytes cultured in normal Petri dishes producing 36.4 and 31.7 pg/cell/h concentrations of urea, respectively. Albumin secretion rate was maximal on day 3 after plating (0.86 pg/cell/h). When using culture medium supplemented with 10% FBS, the metabolic functions of hepatocytes were higher compared to when using normal culture medium. This study demonstrates that human hepatocytes can be isolated from HCV-infected livers via surgical biopsies with high cell yield rate, and their metabolic functions remain stable when cultured in collagen-coated dishes with FBS-rich medium for several days. Base on these results, HCV-infected human hepatocytes can be used in the study of clinical diseases and in the performance of physiological and pharmacological liver experiments in vitro.