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目的探索无血清培养H3N2流感病毒冷适应株的适宜条件。方法以无血清适应的Vero细胞为介质,对流感病毒冷适应株(H3N2)的培养条件进行优化,以得到较优的基础培养基、p H值、TPCK胰酶添加量及其补加时间、病毒接种量和病毒收获时间。结果 H3N2流感病毒冷适应株能够感染无血清适应的Vero细胞;经过优化,病毒培养液适宜p H为7.15~7.64,BSA和TPCK胰酶添加量均为0.8~1.2μg/ml,细胞培养36h接种病毒,接种量MOI为0.05~0.10,于25℃培养168h收获时病毒效价达最高。结论优化了H3N2流感病毒冷适应株无血清培养的适宜条件。
Objective To explore the suitable conditions of serum-free culture of H3N2 influenza virus cold-adapted strain. Methods The culture conditions of influenza virus cold-adapted strain (H3N2) were optimized with serum-free Vero cells to obtain better basic culture medium, p H value, addition of TPCK trypsin and its addition time, Virus inoculation and virus harvest time. Results The H3N2 influenza virus cold-adapted strain could infect Vero cells without serum adaptation. After optimization, the optimum pH for virus culture medium was 7.15-7.64, the addition amount of BSA and TPCK trypsin was 0.8-1.2μg / ml, and the cells were cultured for 36h Virus, inoculation MOI of 0.05 to 0.10, cultured at 25 ℃ 168h virus titers reached the highest price. Conclusion The optimal conditions of serum-free culture of H3N2 influenza virus cold-adapted strain were optimized.