Production of Bovine Herpesvirus-1 Vaccine Strains in MDBK Cells Using BelloCell Bioreactor

Abstract

Aim: To produce bovine herpesvirus-1 (BoHV-1) vaccine strains in MDBK cells using an oscillating bioreactor, BelloCell. Place and Duration of Study: Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar-243122, India, between November 2013 and June 2015. Methodology: MDBK cells were cultured in BelloCell bioreactor seeded with MDBK cells. After 6 days incubation at 37°C under 5% CO2 tension, the MDBK cells were infected with 0.001 multiplicity of infection (MOI) of either wild-type (wt)-BoHV-1 or glycoprotein E deleted (∆gE)-BoHV-1vaccine strains and incubated further for 96 h. To analyse the vaccine potential of the harvests, inactivated wt-BoHV-1 and ∆gE-BoHV-1 were inoculated in guinea pigs. Sera from immunized guinea pigs were collected at 90 days after primary immunization and analysed for virus neutralizing (VN) antibody response specific to wt-BoHV-1. Results: Using BelloCell bioreactor, the MDBK cell density reached to a maximum number of         1.5 x 109 to 2.0 x 109 cells after 6 days cultivation. After infection with wt-BoHV-1 and ∆gE-BoHV-1 strains, the total virus yield at 96 h post-infection was 0.5 x 1011.25 and 0.5 x 1011.83, respectively. There was no significant difference in VN antibody response in sera collected from immunized guinea pigs. Conclusion: This study indicated that BelloCell bioreactor can be used as a simple system for high population density cell culture and large scale production of vaccines.