ST悬浮细胞培养猪伪狂犬病病毒工艺参数的优化

Optimization of Technological Parameters for Culturing PRV in ST Suspension Cells

为优化ST悬浮细胞培养条件及其生产猪伪狂犬病病毒的工艺参数,对影响ST悬浮细胞生长的接种细胞初始密度、培养时间和摇瓶转速等工艺参数进行优化比较,对影响猪伪狂犬病病毒增殖的接种细胞初始密度、感染量和培养时间等条件进行优化。结果显示:接种细胞初始密度1.00×10^(6)cells/mL、摇瓶转速140 r/min、悬浮培养48 h时,细胞数量扩增了4倍且细胞活力高;猪伪狂犬病病毒接种时初始细胞密度2.00×10^(6)cells/mL、感染量MOI=1.0、培养60~72 h,毒价可达10^(9.00)TCID_(50)/mL。结果表明,优化后的培养工艺适用于摇瓶中ST悬浮细胞及猪伪狂犬病病毒的培养。本研究为获得高密度ST悬浮细胞和提高猪伪狂犬病病毒繁殖能力提供了试验依据。

In order to optimize the culture conditions of ST suspension cells and the technological parameters for culturing porcine pseudorabies virus(PRV),the technological parameters including initial density of inoculated cells,culture time and shaking-flask speed that would affect the growth of ST suspension cells were optimized and compared,then the conditions including initial density of inoculated cells,infection amount and culture time that would affect the proliferation of PRV were optimized.The results showed that the number of cells was amplified four times with strong vitality when the initial density of inoculated cells was 1.00×10^(6)cells/mL and the shaking-flask speed was 140 r/min for 4 hours under suspension culture;the initial density was 2.00×10^(6)cells/mL,the infection amount was MOI=1.0 and the virus titer could reach up to 10^(9.00)TCID_(50)/mL for 60 to 72 hours under culture when PRV was inoculated.It was concluded that the optimized parameters were suitable for culture of ST suspension cells and PRV in flask,and an experimental basis was provided by the paper for obtaining ST suspension cells with high density and improving the reproductive ability of PRV.