Vero细胞微载体技术规模化培养轮状病毒

Large Scale Culture of Rotavirus by Vero Cell Microcarrier Technique

目的利用Vero细胞微载体技术规模化培养轮状病毒。方法利用5 L生物反应器进行Vero细胞的微载体培养,待细胞密度达1.5×106个/ml时,以0.05 MOI接种轮状病毒P[2]G3株,于37℃连续培养6 d后收获病毒,期间每天取样观察细胞病变(CPE),并检测病毒感染性滴度。结果在轮状病毒规模化培养的初期,其病毒滴度逐渐升高,至48 h达到最高。在上清中,病毒的滴度为5.5CCID50/ml;在细胞裂解产物中,病毒的滴度为3.75CCID50/ml。结论Vero细胞微载体规模化培养轮状病毒可提高病毒的滴度。

Objective To culture rotavirus in a large scale by using Vero cell microcarrier technique.Methods Culture Vero ceils with microcarrier in a 5 L bioreactor,to which rotavirus strain P[2]G3 was insulated at a MOI of 0.05 when the ceil density reached 1.5 × 106 cells/ml.Incubate the mixture at 37℃ for 6 d and take samples for observation of CPE and determination of infectious titer of virus daily. Results The infectious titer of virus increased gradually at early stage of culture and reached a peak value 48 h after culture. The virus titer in culture supernatant and lysate of cells were 5.5 and 3.75 CCID50/ml respectively. Conclusion The titer of rotavirus after large scale culture by Vero ceil microcarrier technique increased significantly.