无血清全悬浮PK15细胞培养猪圆环病毒2型的研究

The study of suspended PK15 cells culture porcine circovirus type 2 by serum-free medium

关键词:  猪圆环病毒2型  全悬浮  PK15细胞  无血清  连续收获

Key words:  porcine circovirus type 2  suspension  PK15 cells  serum free  continuous harvest

摘要:

为了改进猪圆环病毒2型的培养工艺,驯化了一株可无血清培养的全悬浮PK15细胞用于培养猪圆环病毒2型,并对病毒的敏感性、接毒时间、MOI、收获方法进行了试验。结果表明,该细胞培养猪圆环病毒2型,批次收获时,接毒时细胞密度为0.5×10_⁶/mL,接毒量为0.1 MOI,接毒后72 h后收毒,病毒滴度能达到10_^6.4 TCID₅₀/mL；如果采用连续收获,接毒和带毒传代时均使细胞密度为0.5×10_⁶/mL,接毒量为0.04 MOI,收毒时间为接毒后72 h,连续收获3代,病毒滴度分别为10_⁶ TCID₅₀/mL、10_^6.25 TCID₅₀/mL、10_^6.5 TCID₅₀/mL,病毒滴度和收获液体积较为理想,以期为大规模培养猪圆环病毒2型提供参考。

Abstract:

In order to improve the culture of pocine circovirus type 2, a suspension PK15 cell line that could be cultured by serum-free medium was domesticated to culture pocine circovirus type 2 . The virus sensitivity, the infection time, MOI and harvest methods were investigated. Results shown that after 72 h of infection when using 0.1 MOI of pocine circovirus type 2 to infect the domesticated suspension PK15 cell in the cell density of 0.5×10_⁶ cells per 1 mL, the title of virus could reach to 10_^6.4 TCID₅₀/mL. In addition, when using 0.04 MOI of pocine circovirus type 2 to infect the domesticated suspension PK15, and continuous harvested three times after 72 h of infection, the virus title could reach to 10_⁶ TCID₅₀/mL, 10_^6.25 TCID₅₀/mL, 10_^6.4 TCID₅₀/mL. In conlusion, this study provides a reference for large-scale culture of porcine circovirus type 2.