PK-15细胞在微载体悬浮放大培养及PCV2增殖工艺研究  

Scale-up Process of Microcarrier Suspension Culture of PK-15 Cells and Propagation of PCV2

作　　者：华涛[1,2] 冯磊[1,2] 张雪花[1,2] 唐波[1,2] 常晨[1,2] 刘国阳[1,2] 吴培培[1,2] 陈丽[1,2] 张道华[1,2] 侯继波[1,2] 

机构地区：[1]江苏省农业科学院国家兽用生物制品工程技术研究中心,农业部兽用生物制品工程技术重点实验室,江苏南京210014 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《西南农业学报》2017年第2期474-480,共7页Southwest China Journal of Agricultural Sciences

基　　金：江苏省农业自主创新项目[CX(14)5044];农业部公益性行业(农业)科研专项经费项目(201303046);江苏省农业支撑计划(BE2012370)

摘　　要：为建立在生物反应器内微载体逐级放大培养PK-15细胞和增殖PCV2技术。采用分批式培养方法,研究了PK-15细胞在微载体胰酶消化放大悬浮培养及猪圆环病毒2型(PCV2)增殖工艺。结果表明,5 g/L的微载体和高糖DMEM下,采用4.0×105cells/m L的初始接种密度及培养至第2天进行换液操作工艺可以获得最佳的PK-15细胞生长效能。胰酶消化转移将PK-15细胞从1 L反应器放大至3 L反应器,微载体上细胞贴附均匀、生长旺盛,培养3 d细胞密度可达34.3×105cells/m L。采用感染复数为0.1的接毒比例,细胞接毒后在微载体上传至第3代收获毒液可获得最高的PCV2增殖效价107.25TCID50/m L。为PCV2疫苗的生物反应器规模化生产奠定了基础。In order to develop a technique for scale-up of microcarrier suspension culture of PK-15 cells and propagation of PCV2 in bioreae- tor,batch-wise suspension culture of PK-15 cells on microcarriers （ cytodex 1 ） using scale-up of tripsin digest and propagation of porcine cir- covirus type 2 （PCV2） were investigated. The results showed that the optimal growth efficiency could be achieved by 4.0 × 10^5 cells per 1 mL inoculation on 5 g/L concentration, DMEM medium with high glucose and media replacement after 2-day culture. The highest PCV2 titer was achieved 10^7.25TCIDso/mL at the 3^rd passage of PK-15 cells infected with 0.1 multiple of infection （MOI） of PCV2 The optimal processes achieved in this study laid the experimental foundation for mass production of PCV2 vaccines in bioreactor scale.

关 键 词：PK-15细胞 微载体 消化放大 猪圆环病毒2型 悬浮培养 

分 类 号：Q78[生物学—分子生物学]