In Vitro Infection with Hepatitis B Virus Using Differentiated Human Serum Culture of Huh7.5-NTCP Cells without Requiring Dimethyl Sulfoxide

• Published in: Viruses Vol. 13; no. 1; p. 97
• Main Authors: Le, Connie, Sirajee, Reshma, Steenbergen, Rineke, Joyce, Michael A, Addison, William R, Tyrrell, D Lorne
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 12-01-2021; MDPI AG
• Subjects: Biomarkers; Cell Line; Cells, Cultured; differentiated Huh7.5-NTCP human serum culture; dimethyl sulfoxide (DMSO); Dimethyl Sulfoxide - pharmacology; Gene Expression; Genetic Vectors - genetics; Hepatitis B - virology; hepatitis B virus (HBV); Hepatitis B virus - drug effects; Hepatitis B virus - physiology; Hepatocytes - drug effects; Hepatocytes - metabolism; Hepatocytes - virology; hepatoma cell culture; Humans; Organic Anion Transporters, Sodium-Dependent - genetics; sodium taurocholate co-transporting polypeptide (NTCP); Symporters - genetics; Virus Internalization - drug effects; Virus Replication - drug effects; dimethyl sulfoxide (DMSO); hepatoma cell culture; hepatitis B virus (HBV); sodium taurocholate co-transporting polypeptide (NTCP); differentiated Huh7.5-NTCP human serum culture
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v13010097

An estimated two billion people worldwide have been infected with hepatitis B virus (HBV). Despite the high infectivity of HBV in vivo, a lack of easily infectable in vitro culture systems hinders studies of HBV. Overexpression of the sodium taurocholate co-transporting polypeptide (NTCP) bile acid transporter in hepatoma cells improved infection efficiency. We report here a hepatoma cell culture system that does not require dimethyl sulfoxide (DMSO) for HBV infection. We overexpressed NTCP in Huh7.5 cells and allowed these cells to differentiate in a medium supplemented with human serum (HS) instead of fetal bovine serum (FBS). We show that human serum culture enhanced HBV infection in Huh7.5-NTCP cells, e.g., in HS cultures, HBV pgRNA levels were increased by as much as 200-fold in comparison with FBS cultures and 19-fold in comparison with FBS+DMSO cultures. Human serum culture increased levels of hepatocyte differentiation markers, such as albumin secretion, in Huh7.5-NTCP cells to similar levels found in primary human hepatocytes. N-glycosylation of NTCP induced by culture in human serum may contribute to viral entry. Our study demonstrates an in vitro HBV infection of Huh7.5-NTCP cells without the use of potentially toxic DMSO.