Marc-145细胞微载体悬浮培养及PRRSV增殖工艺的建立

采用分批式培养方法研究了Marc-145细胞微载体悬浮培养及猪繁殖与呼吸综合征病毒(PRRSV)增殖工艺。结果表明,在3 g/L的微载体用量下,采用1 ml 3×105个细胞的初始接种密度及培养至第3 d进行换液操作工艺可以获得最佳的Marc-145细胞生长效能。采用感染复数为0.05的接毒比例及接毒后48 h收获毒液可获得最高的PRRSV增殖效价。在5 L反应器中重复验证上述工艺,获得较为稳定的试验结果,最大细胞密度均可高于1 ml 2×106个细胞,PRRSV的增殖效价均高于108.0TCID50/ml。为猪繁殖与呼吸综合征病毒疫苗的生物反应器规模化生产奠定了基础。

Microcarrier suspension culture of Marc-145 cells and propagation of PRRSV

Batch-wise suspension culture of Marc-145 cells on microcarriers(cytodex 1) and propagation of porcine reproductive and respiratory syndrome virus(PRRSV) were investigated.The optimal growth efficiency could be achieved by 3×105 cells per 1 ml inoculation on 3 g/L microcarriers concentration and media replacement after 3-d culture.The highest PRRSV titer was achieved with 0.05 multiple of infection(MOI) on Marc-145 cells at 48 h post infection.All conditions for Marc-145 cells microcarriers suspension culture and PRRSV propagation mentioned above were confirmed in 5 L bioreactor repeatedly,resulting in the highest cell density above 2×106 cells per 1 ml and PRRSV titer above 108.0 TCID50/ml.The optimal processes achieved in this study laid the experimental foundation for mass production of PRRS vaccines in bioreactor scale.