微生物学免疫学进展
微生物學免疫學進展
미생물학면역학진전
PROGRESS IN MICROBIOLOGY AND IMMUNOLOGY
2014年
1期
25-30
,共6页
安红%张海红%王名强%韩平%余黎%周旭
安紅%張海紅%王名彊%韓平%餘黎%週旭
안홍%장해홍%왕명강%한평%여려%주욱
轮状病毒%Vero细胞%细胞工厂%病毒滴度
輪狀病毒%Vero細胞%細胞工廠%病毒滴度
륜상병독%Vero세포%세포공엄%병독적도
Rotavirus%Vero cells%Cell factory%Virus titer
目的:以细胞工厂代替转瓶培养轮状病毒基因重配株Ls的可行性研究。方法采用细胞工厂与相应的转瓶培养工艺作对比,比较两种容器内细胞生长状态与病毒收获液滴度,并对细胞工厂培养条件进行了优化。结果以相同浓度接种细胞时,细胞工厂4 d长成单层,转瓶却需要7 d,经细胞仪计数后单位面积内细胞密度相当;以相同MOI接种病毒后,转瓶内的病毒于第7天病毒滴度达到峰值,细胞已完全脱落;细胞工厂于第3天病毒滴度达到峰值,并实现了3次收获。细胞工厂每次收获的病毒液滴度都稳定在一定范围,与转瓶相当。另外,细胞工厂培养条件优化结果表明,Vero细胞最佳接种浓度为3.0×104细胞/cm2,接种病毒的最适MOI为0.02~0.04。结论使用细胞工厂培养Ls株病毒不仅提高了效率,而且减少了培养空间,可替代转瓶规模化生产轮状病毒疫苗。
目的:以細胞工廠代替轉瓶培養輪狀病毒基因重配株Ls的可行性研究。方法採用細胞工廠與相應的轉瓶培養工藝作對比,比較兩種容器內細胞生長狀態與病毒收穫液滴度,併對細胞工廠培養條件進行瞭優化。結果以相同濃度接種細胞時,細胞工廠4 d長成單層,轉瓶卻需要7 d,經細胞儀計數後單位麵積內細胞密度相噹;以相同MOI接種病毒後,轉瓶內的病毒于第7天病毒滴度達到峰值,細胞已完全脫落;細胞工廠于第3天病毒滴度達到峰值,併實現瞭3次收穫。細胞工廠每次收穫的病毒液滴度都穩定在一定範圍,與轉瓶相噹。另外,細胞工廠培養條件優化結果錶明,Vero細胞最佳接種濃度為3.0×104細胞/cm2,接種病毒的最適MOI為0.02~0.04。結論使用細胞工廠培養Ls株病毒不僅提高瞭效率,而且減少瞭培養空間,可替代轉瓶規模化生產輪狀病毒疫苗。
목적:이세포공엄대체전병배양륜상병독기인중배주Ls적가행성연구。방법채용세포공엄여상응적전병배양공예작대비,비교량충용기내세포생장상태여병독수획액적도,병대세포공엄배양조건진행료우화。결과이상동농도접충세포시,세포공엄4 d장성단층,전병각수요7 d,경세포의계수후단위면적내세포밀도상당;이상동MOI접충병독후,전병내적병독우제7천병독적도체도봉치,세포이완전탈락;세포공엄우제3천병독적도체도봉치,병실현료3차수획。세포공엄매차수획적병독액적도도은정재일정범위,여전병상당。령외,세포공엄배양조건우화결과표명,Vero세포최가접충농도위3.0×104세포/cm2,접충병독적최괄MOI위0.02~0.04。결론사용세포공엄배양Ls주병독불부제고료효솔,이차감소료배양공간,가체대전병규모화생산륜상병독역묘。
Objective To estimate the possibility of culturing the reassortant rotaviruses strain Ls on a cell factory instead of a conventional roller bottle .Methods The growths of cells and titers of harvested virus cultured by the two methods were compared.Furthermore, the condition for culture on the cell factory was optimized .Results The results indicate that the monolayer is formed on the surface of the cell factory within 4 d of cultivation , but on the surface of the roller bottle within 7 d, when the Vero cells are inoculated at the same cell concentration .We found that the cell density of two contain-ers is equal by using the cell counter .The titer of viruses in roller bottle reaches a peak value at 7th day, afterwards cells have completely exfoliated;Those cultured on the cell factory achieve the highest titer of viruses at the 3rd day, and could be harvested for 3 times.The titers of each virus harvest are stable within a certain range on the cell factory , which are equal to those of the roller culture.The optimal inoculation concentration and MOI are 3.0×104 cells/cm2 and 0.02~0. 04, respectively.Conclusion The efficiency of culturing the rotaviruses strain Ls on the cell factory is not only increased , but the culture space is decreased , which shows that the cell factory could be used in a large-scale production of rotavirus vaccine instead of the roller bottle .