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In Vitro Infection with Hepatitis B Virus Using Differentiated Human Serum Culture of Huh7.5-NTCP Cells without Requiring Dimethyl Sulfoxide
| Content Provider | MDPI |
|---|---|
| Author | Le, Connie Sirajee, Reshma Steenbergen, Rineke Joyce, Michael A. Addison, William R. Tyrrell, D. Lorne |
| Copyright Year | 2021 |
| Description | 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. |
| Starting Page | 97 |
| e-ISSN | 19994915 |
| DOI | 10.3390/v13010097 |
| Journal | Viruses |
| Issue Number | 1 |
| Volume Number | 13 |
| Language | English |
| Publisher | MDPI |
| Publisher Date | 2021-01-12 |
| Access Restriction | Open |
| Subject Keyword | Viruses Virology Hepatitis B Virus (hbv) Hepatoma Cell Culture Sodium Taurocholate Co-transporting Polypeptide (ntcp) Differentiated Huh7.5-ntcp Human Serum Culture Dimethyl Sulfoxide (dmso) |
| Content Type | Text |
| Resource Type | Article |
