High vero cell density and rabies virus proliferation on fibracel disks versus cytodex-1 in spinner flask.

ABSTRACT

To achieve higher titer of rabies virus higher density of host cells will need. In this study, capability of FibraCel disks packed in 500 mL spinner basket versus Cytodex-1 in 500 mL spinner flask was investigated for propagation of Vero cells and PV rabies virus proliferation. Minimal Essential Medium (MEM) + 10% Foetal Calf Serum (FCS) and Virus Production- Serum Free Medium (VP-SFM) +4 mM L-glutamine were used in growth phase and MEM+ 0.2% Bovine Serum Albumin (BSA) and VP-SFM were used in virus production phase. Adapted Vero cells grown in VP-SFM were used in all SFM experiments while batch and stepwise perfusion modes were applied and compared in growth stage. The highest Vero cell density were achieved in the trials with 10 g FibraCel disk in stepwise perfusion mode equal to 6.12 x 10(6) and 5.87 x 10(6) cells mL(-1) in MEM and VP-SFM, respectively while with 2.73 g Cytodex-1 lower density equal to 4.2 x 10(6) and 4.0 x 10(6) cells mL(-1) were achieved. The highest titer of rabies virus and overall virus production rate were resulted in VP-SFM and on 10 g disks equal to 2.9 x 10(7) Fluorescent Focus Unit (FFU) mL(-1) and 0.14 FFU/Cell/h, respectively versus 1.7 x 10(7) FFU mL(-1) and 0.08 FFU/cell/h on cytodex-1 in similar conditions. The second harvest of virus was also satisfactory in experiment with 10 g disks (1.7 x 10(7) FFU mL(-1)) in compare to Cytodex-1 (0.51 x 10(7) FFU mL(-1)). An equal surface area at 6600 and 12000 cm(-2) were provided in all comparable trials with seeding density of 12.5 x 10(3) cells cm(-2). Adapted Vero cells grown in VP-SFM were used in all SFM experiments while batch and stepwise perfusion modes were applied and compared in growth stage.