Seminar: Dr. Rineke Steenbergen

Drastically increasing hepatitis C yield by growth in human serum, instead of fetal bovine serum

Date:
23 September 2014 00:00 hrs.
Location:
Location see remarks
Title:
Drastically increasing hepatitis C yield by growth in human serum, instead of fetal bovine serum
Speaker(s):

Dr. Rineke Steenbergen, University of Alberta, Canada

Host(s):

Dr. Bas Dutihl, Dept. of CMBI, Radboudumc

23-09-2014 00:00:00Europe/AmsterdamDrastically increasing hepatitis C yield by growth in human serum, instead of fetal bovine serum Location see remarksRimlsrimls@radboudumc.nl

Remarks / more information:

SteenbergenDr. Rineke Steenbergen is a senior research scientist at the Li Ka Shing Institute of Virology and the Applied Virology Institute, both located at the University of Alberta in Edmonton, Canada. She has investigated knockout mice with defects in lipid biosynthesis, and the role of lipoproteins in the life cycle of Hepatitis C Virus. Her current research interests focus on the role lipid of metabolism in the life cycle of Hepatitis A, B and C viruses, and the development of a vaccine against Hepatitis C Virus.

When a human hepatocellular carcinoma cells (Huh7.5) are cultured in media containing human serum (HS), instead of the standard fetal bovine serum (FBS), the cells undergo growth arrest, and differentiate into a hepatocyte like cell type. These cells show increased expression of hepatocyte specific markers and become polarized. Additionally, hepatocyte specific functions, that were absent under FBS conditions, are restored in HS cultured cells, including very low density lipoprotein secretion. Microarray analysis revealed that gene expression of approximately 22-30% of the genes had significantly changed. Pathways enrichment analysis showed that xenobiotics metabolism by cytochrome P450, another liver specific function, was among the pathways that were significantly increased.

We showed that culturing Huh7.5 cells in HS resulted in a 1000-fold increase in HCV production, and the virus that is produced in HS more closely resembles the viruses that circulate in the blood of patients. These cells also become susceptible to Hepatitis B virus (HBV) infection.

Overall, our data show that Huh7.5 cells that are cultured in HS containing media become more hepatocyte-like, in morphology as well as in function.

 



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