微载体悬浮培养ST细胞增殖猪瘟兔化弱毒的工艺研究

Propagation of classical swine fever virus HCLV strain by microcarrier suspension culture of ST cell

关键词:  微载体悬浮培养、ST细胞、猪瘟兔化弱毒

Key words:  microcarrier suspension culture, ST cells, HCLV

摘要:

本研究从细胞的接种密度、病毒接毒量、培养基三个方面进行了研究和优化，并进行了放大培养，建立猪瘟病毒的微载体悬浮培养工艺，确定微载体使用浓度3g/L，细胞接种密度为每个微载体15个细胞，病毒接毒量0.05MOI,采用DMEM/F12培养基进行培养和细胞消化瓶批式消化分散细胞，培养的细胞可以完成生物反应器10L到50L的放大，培养病毒可使含量达到7.6lgTCID50/ml、不低于400万RID/ml，工艺稳定可靠，为猪瘟疫苗工业化大规模微载体悬浮培养奠定了基础。

Abstract:

In order to develop the technology for microcarrier suspension culture of ST cells and propagation of classical swine fever virus HCLV strain in bioreactor, the cell density,amount of virus inoculation, the culture medium and Scale-up Process were studied and optimized. Results showed that the virus titerofHCLV can reached to 7.6lgTCID50/ml and more than four million RID/ml, when using the initial seeding density of 15 cells per microcarrier under the dosage of 3g /L microcarrier,DMEM/F12 medium and infected with 0.05 multiple of infection ( MOI) of HCLV. Flask batch digestion can of scale-up the cultures from 10L to 50L. All together this suspension culture process is stable and reliable, whichlaid the foundation for large scale microcarrier suspension culture of swine fever vaccine.