Application of a serum-free medium for the growth of Vero cells and the production of reovirus.
Two strains of reovirus (serotype 1 Lang/TIL and serotype 3 Dearing/T3D) were propagated in Vero cells grown in stationary or agitated cultures in a serum-free medium, M-VSFM. Solid microcarriers (Cytodex-1) were used to support cell growth in agitated cultures with a normal doubling time of 25 h. Cell yields of 1 x 10(6) cells/mL were obtained from an inoculum of 2 x 10(5) cells/mL in 4 days in microcarrier cultures. The growth profile and cell yield was not significantly different from serum-supplemented cultures. The virus titer increased by 3-4 orders of magnitude over a culture period of 150 h. The maximum virus titer in stationary cultures reached >1 x 10(9) pfu/mL for both strains of reovirus in M-VSFM. M-VSFM also supported high viral yields in microcarrier cultures. Both the specific productivity and final viral yield was higher in M-VSFM than serum-supplemented cultures. The high viral productivity suggests that this is a suitable system for the production of reovirus as an oncolytic agent for human therapeutic use.
Butler
M
M
Departments of Microbiology and Medical Microbiology, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada. butler@cc.umanitoba.ca
Burgener
A
A
Patrick
M
M
Berry
M
M
Moffatt
D
D
Huzel
N
N
Barnabé
N
N
Coombs
K
K
D000818:Animals; D002455:Cell Division; D002522:Chlorocebus aethiops; D016895:Culture Media, Serum-Free; D012087:Reoviridae / Q000502:physiology*; D014709:Vero Cells; D010948:Viral Plaque Assay; D014779:Virus Replication
D016895:Culture Media, Serum-Free