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

L. S. Rajan, A. Kamboj, B. R. Prusty, T. K. Palai, L. Ananthakrishna, M. Saini, P. Gupta
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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% CO 2 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 Original Research Article Rajan et al.; BBJ, 12(1): 1-7, 2016; Article no.BBJ.20401 2 1.5 x 10 to 2.0 x 10 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 10 and 0.5 x 10, 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.
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利用BelloCell生物反应器在MDBK细胞中生产牛疱疹病毒-1疫苗株
目的:利用BelloCell振荡生物反应器在MDBK细胞中生产牛疱疹病毒-1 (BoHV-1)疫苗株。学习地点和时间:2013年11月至2015年6月,印度兽医研究所兽医生物技术部门,Izatnagar-243122,印度。方法:MDBK细胞在接种MDBK细胞的BelloCell生物反应器中培养。在37°C、5% CO 2压力下孵育6天后,MDBK细胞被0.001感染倍数(MOI)的野生型(wt)-BoHV-1或糖蛋白E缺失(∆gE)-BoHV-1疫苗株感染,并进一步孵育96 h。为了分析疫苗潜力,将灭活的wt-BoHV-1和∆gE-BoHV-1接种于豚鼠。在初次免疫后90天收集免疫豚鼠血清,分析wt-BoHV-1特异性病毒中和(VN)抗体反应。结果:使用BelloCell生物反应器,MDBK细胞密度达到Rajan等人的最大数量;植物学报,12(1):1-7,2016;文章no.BBJ。20401 2 1.5 × 10至2.0 × 10细胞培养6天后。wt-BoHV-1和∆gE-BoHV-1感染后96 h总病毒产量分别为0.5 × 10和0.5 × 10。免疫豚鼠血清中VN抗体反应无显著差异。结论:本研究表明,BelloCell生物反应器可作为高密度细胞培养和疫苗大规模生产的简单系统。
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