Potential conflict of interest: Nothing to declare.
Human serum leads to differentiation of human hepatoma cells, restoration of very-low-density lipoprotein secretion, and a 1000-fold increase in HCV Japanese fulminant hepatitis type 1 titers
Article first published online: 17 OCT 2013
© 2013 by the American Association for the Study of Liver Diseases
Volume 58, Issue 6, pages 1907–1917, December 2013
How to Cite
Steenbergen, R. H.G., Joyce, M. A., Thomas, B. S., Jones, D., Law, J., Russell, R., Houghton, M. and Tyrrell, D. L. (2013), Human serum leads to differentiation of human hepatoma cells, restoration of very-low-density lipoprotein secretion, and a 1000-fold increase in HCV Japanese fulminant hepatitis type 1 titers. Hepatology, 58: 1907–1917. doi: 10.1002/hep.26566
The authors thank Dr. Rice for the kind gift of Huh7.5 cells and Dr. Wakita for the JFH-1 construct. This research was funded by the Li Ka Shing Foundation, Canadian Institutes for Health Research, and Alberta Advanced Education and Technology (AAET).
- Issue published online: 26 NOV 2013
- Article first published online: 17 OCT 2013
- Accepted manuscript online: 14 JUN 2013 09:41AM EST
- Manuscript Accepted: 27 MAY 2013
- Manuscript Received: 30 JAN 2013
In this study, we differentiated the human hepatoma cell line Huh7.5 by supplementing tissue culture media with human serum (HS) and examined the production of hepatitis C virus (HCV) by these cells. We compared the standard tissue culture protocol, using media supplemented with 10% fetal bovine serum (FBS), to media supplemented with 2% HS. Cells cultured in HS undergo rapid growth arrest, have a hepatocyte-like morphology, and increase the expression of hepatocyte differentiation markers. In addition, expression of cell adhesion proteins claudin-1, occludin, and e-cadherin are also increased. The lipid droplet content of these cells is highly increased, as are key lipid metabolism regulators liver X receptor alpha, peroxisome proliferator-activated receptor (PPAR)-α, and PPAR-γ. Very-low-density lipoprotein secretion, which is absent in FBS-grown cells, is restored in Huh7.5 cells that are cultured in HS. All these factors have been implicated in the life cycle of HCV. We show that viral production of Japanese fulminant hepatitis type 1 increases 1,000-fold when cells are grown in HS, compared to standard FBS culture conditions. The virus produced under these conditions is associated with apolipoprotein B, has a lower density, higher specific infectivity, and has a longer half-life than virus produced in media supplemented with FBS. 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)