Monday, July 30, 2007
P32

Microcarrier Culture for Dengue Vaccine Development: Comparing Serum-Containing and Serum-Free Media, and Virus Genomic Stability in Vero and MRC-5 Cells

Suh-Chin Wu1, Chia-Chyi Liu1, Shiang-Chi Lee1, Mike Butler2, and Ching-Juh Lai3. (1) Institute of Biotechnology and Department of Life Science, National Tsing Hua University, 101, Section 2, Guangfu Road, Hsinchu, 30013, Taiwan, (2) Department of Microbiology, University of Manitoba, Winnipeg, Canada, (3) Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892

This study investigated the production processes of four-serotype and infectious clone dengue (DEN) viruses from Vero and MRC-5 cells grown on Cytodex 1 microcarriers.  Four-serotype strains of DEN-1 (HAWAII strain), DEN-2 (NGC strain), DEN-3 (H-87 strain), DEN-4 (H-241 strain), and the infectious clone 2A-derived DEN-4 virus were used to investigate the replication kinetics and genomic stability of DEN viruses produced in serum-free and serum-containing microcarrier cultures.  Serum-free microcarrier cultures were found to yield significantly higher titers than serum-containing cultures to produce four-serotype DEN viruses particularly in MRC-5 cells.  Serum-free microcarrier cultures were also effective to propagate the infectious clone 2A-derived DEN-4 viruses in Vero and MRC-5 cells, but the maximum titers in MRC-5 cells were one-log lower than Vero cells.  However, genomic sequences of the infectious clone 2A-derived DEN-4 viruses produced in serum-free and serum-containing cultures indicated that MRC-5 cells were more efficient to maintain the genomic stability of DEN virus propagation in cell cultures.  Taken together, MRC-5 cells are more favorable than Vero cells for DEN virus production.  This result provides valuable information on cell culture systems for live-attenuated DEN vaccine development.

This work was supported by the National Health Research Institutes, Taiwan (NHRI-EX96-9534SI)


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