The aim of this work was to establish a process to produce influenza virus in Vero cell culture using single-use equipment. The end result would be a fast and simple process enabling scale-up and adaptation to industrial production.
The following parameters were evaluated: medium composition, cell detachment, medium supplements, agitation conditions, and infection conditions. Different combinations of media and proteases were evaluated for cell growth in static culture. As part of optimizing the conditions for cell growth in WAVE Bioreactor system, the effects of a number of supplements and different rocking conditions were evaluated. Next, in order to find the optimal infection parameters, both trypsin concentration and different virus stock dilutions were evaluated. Finally, the robustness and reproducibility of the entire workflow for virus propagation in Vero cells using microcarriers and WAVE Bioreactor system was investigated.
The data show that a robust process using WAVE Bioreactor system for propagating influenza virus in Vero cells attached to Cytodex 1 microcarriers has been developed. WAVE Bioreactor 20/50 system using a single-use Cellbag* bioreactor (Fig 1) offers easy set-up with no cleaning required, making it a fast and convenient alternative to stainless steel bioreactors.