Optimization of mammalian reovirus T3D growth in L929 cells

Abstract

Although the demand for high scale production of mammalian orthoreoviruses (T3D in particular) is mounting to advance the research studies and clinical trials associated with their oncolytic capacity, very few reported studies have tried to understand the factors that affect the its production and subsequently optimize them to maximize the virus yield in cell culture systems. For this study, we manipulated several growth parameters (multiplicity of infection, cell density, cell feeding, media pH, and flask size) sequentially to select optimal conditions. Manipulation of cell density, whether cells were fed or not, and flask size all led to moderate changes in progeny virus titer. Altered media pH led to dramatic (more than 100 fold) virus replication changes. We conclude that cell physiological status has a key impact on infection progression and virus production. Optimal parameters consisted of infection at MOI of 0.1, cell density at infection of 95%, media re-feeding, media pH 7.0-8.5, and the smaller the flask size the better.