Vero细胞和脊髓灰质炎病毒在无血清条件下的培养探究

Investigation on cultivation of Vero cell and poliovirus under serum-free condition

目的探索Vero细胞和脊髓灰质炎病毒在无血清条件下的最佳培养条件,为无血清培养Vero细胞生产脊髓灰质炎疫苗奠定基础。方法选择直接适应(直降组)和序贯适应(驯化组)两种无血清培养方法,观察Vero细胞和脊髓灰质炎病毒在无血清条件下的生长情况,并检测脊髓灰质炎病毒及其病毒滴度。结果 Vero细胞在两种无血清条件下均生长良好,其中驯化组细胞生长速度更接近对照组。以脊髓灰质炎病毒Sabin株Ⅰ型分别感染直降组、驯化组和对照组细胞的病毒滴度平均值分别为8.94、8.81和8.94 LgCCID50/mL;Ⅱ型病毒滴度平均值分别为8.84、8.25和7.94 LgCCID50/mL;Ⅲ型病毒滴度平均值分别为8.91、8.57和8.63 LgCCID50/mL;且3组的变异系数(CV)均小于10%。结论 Vero细胞在无血清条件下生长良好,无血清培养的Vero细胞可用作脊髓灰质炎疫苗生产的基质。

Objective To explore the optimal condition for serum-free cultivation of Vero cell and poliovirus.Methods Two methods were carried out for cultivation of Vero cell by using serum-free medium including a direct adaptation （ direct adaption group ） and a sequential adaptation （ acclimatization group ） .Vero cells and poliovirus were cultured and observed in serum-free conditions , and poliovirus titers were detected too .Results The growth status of Vero cell was in good con-dition cultured in the two kinds of serum free media , and the cell growth rate in acclimation group is approximatelly to reach that in control group .The mean value of virus titers were 8.94 lgCCID50/mL, 8.81 lgCCID50/mL and 8.94 lgCCID50/mL respectively in Vero cell infected with poliovirus typeⅠSabin strain for direct adaption group , acclimatization group and con-trol group.The mean value of virus titers were 8.84 lgCCID50 mL, 8.25 lgCCID50/mL and 7.94 lgCCID50/mL respectively in Vero cell infected with poliovirus typeⅡSabin strain in three groups .The mean value of virus titers was 8.91 lgC-CID50 mL, 8.57 lgCCID50/mL and 8.63 lgCCID50/mL respectively in Vero cell infected with poliovirus typeⅢSabin strain in three groups .The coefficient of variation （ CV） values for all three groups were less than 10%.Conclusion The cul-ture of Vero cell in serum-free condition is successful for the proliferation of poliovirus used as a substrate in production of vaccines .