国产与进口无血清培养基和重组胰蛋白酶规模化培养Vero细胞效果的比较

Comparison of large-scale culture of Vero cells with serum-free media and recombinant trypsins from domestic and import venders

目的比较国产物料与进口物料对Vero细胞的培养效果及消化效果,以判断国产物料VirusPro?VP无血清培养基(serum-free medium,SFM)(干粉)和Trpzyme重组胰蛋白酶是否适用于规模化Vero细胞培养。方法选取国产物料VirusPro?VP SFM(干粉)和Trpzyme作为实验组,选取进口物料粉剂VP SFM和TryPLe Select作为对照组,在T瓶、10层细胞工厂(10-layer cell factory,CF10)和CF40连续规模化生产中,对特定代次、特定传代规模分别进行独立样本t检验,比较总消化时长、细胞收获密度和细胞存活率。结果各代次中,国产物料组与进口物料组的细胞生长状态均相似且良好。T瓶、CF10和CF40传代的独立样本t检验结果显示,虽然国产物料组的T瓶、CF10、CF40总消化时长(17.33、19.17、19.17 min)均分别大于进口物料组(9.17、16.33、17.42 min),但仅T瓶总消化时长差异具有统计学意义(t=-5.58,P<0.05)。两组间细胞收获密度及细胞存活率的差异均不具有统计学意义(P>0.05),T瓶传代阶段的细胞收获密度(进口物料组:3.03×10^(6)ml^(-1);国产物料组:3.11×10^(6)ml^(-1))均高于CF10(进口物料组:1.22×10^(6)ml^(-1);国产物料组:1.13×10^(6)ml^(-1))和CF40(进口物料组:9.90×10^(5)ml^(-1);国产物料组:8.73×10^(5)ml^(-1))。结论国产物料VirusPro?VP SFM(干粉)、Trpzyme均符合规模化Vero细胞培养所需。

Objective To compare the culture effect and cell dispersion effect between domestic and imported materials on Vero cells to determine whether domestic materials VirusPro®VP serum free medium(SFM)(dry powder)and Trpzyme recombinant trypsin are suitable for large scale culture of Vero cells.Methods Vero cells were sequentially cultured in T flask and cell factory with domestic materials VirusPro®VP SFM(dry powder)and Trpzyme as test group,and imported materials powder VP SFM and TryPLe Select as control group.Independent-sample t test was performed in T flask,10-layer cell factory(CF10)and 40-layer cell factory(CF40),respectively,to compare period of cell dispersion,cell harvest density and cell viability.Results The cell growth statuses in domestic material group and imported material group were similar and good.The results of independent-sample t test of T flask,CF10 and CF40 showed that,although the total period of cell dispersion in domestic material group(T flask:17.33 min,CF10:19.17 min,CF40:19.17 min)was longer than that in imported material group(T flask:9.17 min,CF10:16.33 min,CF40:17.42 min),there was only a statistically significant difference in the total period cell dispersion between the two groups in T flask(t=-5.58,P<0.05).The cell harvest density and cell viability of the two groups were not statistically significant(P>0.05),and the cell harvest density of T flask(imported material group:3.03×10^(6)ml^(-1);domestic material group:3.11×10^(6)ml^(-1))was higher than those of cell factories(CF10 in imported material group:1.22×10^(6)ml^(-1);CF10 in domestic material group:1.13×10^(6)ml^(-1);CF40 in imported material group:9.90×10^(5)ml^(-1);CF40 in domestic material group:8.73×10^(5)ml^(-1)).Conclusion The domestic VirusPro®VP SFM(dry powder)and Trpzyme are both suitable for large scale culture of Vero cell.