Conclusion: We describe a convenient, cost-effective method to produce hepatocyte-like

cells, which produce large amounts of virus that more closely resemble HCV present in serum of infected patients. (Hepatology 2013; 58:1907–1917) Hepatitis C virus (HCV) is an enveloped, positive-strand RNA virus of the family of Flaviviridae that causes acute and chronic hepatitis. HCV can cause cirrhosis, hepatocellular carcinoma, and steatosis in infected individuals. Replicon systems, both subgenomic and full length, and the Japanese fulminant hepatitis type 1 (JFH-1) tissue culture infection models have yielded important insight into the HCV life cycle. Most of these models make use of HuH-7 or HuH-7-derived cells, such as Huh7.5. HuH-7 or HuH-7-derived Hippo pathway inhibitor cells have many advantages for the in vitro study of HCV: they are readily available and rapidly dividing, and therefore enable large-scale experiments. However, these systems do not necessarily accurately represent events that occur during a natural HCV infection in vivo, because hepatocytes are normally nondividing and fully differentiated. check details To circumvent this, dimethyl sulfoxide (DMSO) has been used to differentiate HuH-7 cells,[1] which resulted in increased expression of hepatocyte-specific genes. These differentiated, growth arrested cells can be infected using

HCV JFH-1 and produce viral titers that are comparable to those in dividing cells.[1] Freshly isolated primary human hepatocytes are a more representative in vitro model to study HCV infectivity. However, viral titers produced in these cells are low, and experiments longer than a few days

require coculture with other cell types.[2, 3] Additionally, primary hepatocytes exhibit large interdonor variability, are often cost prohibitive, and have limited availability. Thus, they are generally not Ponatinib concentration suitable for large-scale experiments. We have previously shown that infection in chimeric mice is not reliably achieved until the humanization of the liver is nearly complete.[4] We postulated that infection with HCV was not only dependent on the presence of human hepatocytes, but also on human factors in serum of mice that have to reach critical levels to support HCV infection. Indeed, we found a good correlation between successful infection and the humanization of lipoprotein profiles in mouse serum.[4] In this study, we extended this observation to Huh7.5 cells in culture and investigated whether the presence of HS in tissue culture is advantageous to infection and viral production. To this end, we compared the “standard” tissue culture protocol, using media containing 10% fetal bovine serum (FBS), to the use of 2% human serum (HS).