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Pakistan Journal of Biological Sciences

Year: 2011 | Volume: 14 | Issue: 7 | Page No.: 441-448
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Authors

S. Mehdi Hassanzadeh

Ali Zavareh

M. Ali Shokrgozar

Ali Ramezani

Ahmad Fayaz

Keywords

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 6.12x106 and 5.87x106 cells mL-1 in MEM and VP-SFM, respectively while with 2.73 g Cytodex-1 lower density equal to 4.2x106 and 4.0x106 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 2.9x107 Fluorescent Focus Unit (FFU) mL-1 and 0.14 FFU/Cell/h, respectively versus 1.7x107 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 (1.7x107 FFU mL-1) in compare to Cytodex-1 (0.51x107 FFU mL-1). An equal surface area at 6600 and 12000 cm-2 were provided in all comparable trials with seeding density of 12.5x103cells 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.
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