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Production of Choristoneura fumiferana Nucleopolyhedrovirus in C. fumiferana (CF-2C1 Cells) in a 3 Litre Bioreactor Using Serum-free Medium
Authors:J Meghrous  A Kamen  S R Palli  S S Sohi  G F Caputo  S C Bedard
Institution:1. Animal Cell Technology and Downstream Processing Group, Biotechnology Research Institute , National Research Council Canada , 6100 Royalmount Avenue, Montreal , Quebec , Canada , H4P 2R2;2. Canadian Forest Service , Great Lakes Forestry Centre , 1219 Queen Street East, P.O. Box 490, Sault Ste. Marie, Ontario , Canada , P6A 5M7;3. Animal Cell Technology and Downstream Processing Group, Biotechnology Research Institute , National Research Council Canada , 6100 Royalmount Avenue, Montreal , Quebec , Canada , H4P 2R2;4. Canadian Forest Service , Great Lakes Forestry Centre , 1219 Queen Street East, P.O. Box 490, Sault Ste. Marie, Ontario , Canada , P6A 5M7
Abstract:The purpose of this study was to develop a cell culture process in a bioreactor for the production of a viral insecticide for the spruce budworm, Choristoneura fumiferana . Several cell lines were tested for their growth in serum-free medium suspension cultures. One cell line, CF-124T-2C1 (CF-2C1), was successfully adapted to grow in suspension cultures in SFM. Serum-free Ex-Cell 405 medium produced a much higher cell density (6.3 x 10 6 cells ml -1 ) than the Grace's medium supplemented with 10% fetal bovine serum (2.5 x 10 6 cells ml -1 ). Also, a higher yield of virus was obtained in the former medium. Ex-Cell 405, was used to study the growth of CF-2C1 cells and the production of C. fumiferana nucleopolyhedrovirus (CfMNPV) in a 3 l bioreactor. Under these conditions, a specific growth rate ( μ) of 0.027 h -1 was obtained during the exponential growth phase, and the specific carbon dioxide evolution rate, as determined by on-line measurement, was 0.9 x 10 -16 mol cell -1 s -1 and 1.78 x 10 -16 mol cell -1 s -1 during growth and infection phases, respectively. Virus production in bioreactor cultures infected at 1.3 x 10 6 cells ml -1 was consistently lower than that obtained in Erlenmeyer shake flasks. Only 26% of the cells were infected in the bioreactor compared to 44% in the shake flasks. However, a higher yield of occluded virus was obtained in the bioreactor cultures than in shake flasks. The production of occlusion bodies (OB) achieved in bioreactor cultures was 2 x 10 6 OB ml -1 .
Keywords:Cfmnpv  Bioreactor  Biopesticide  Choristoneura  Fumiferana
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