摘要
目的用细胞工厂代替转瓶培养沙鼠肾原代细胞,制备肾综合征出血热(HFRS)疫苗,减少原代沙鼠用量,提高产量。方法在细胞工厂内培养沙鼠肾原代细胞,以15L转瓶培养作对照,比较两种容器内细胞生长及收获的病毒滴度和抗原含量;病毒液经超滤浓缩后,采用Sepharose-4FF介质色谱纯化,并进行各项检定。结果细胞工厂只需接种转瓶培养所需1/3量的沙鼠肾细胞,就能在同期与转瓶细胞数量相当。接种病毒后,转瓶中的细胞在收获第5次后已脱落(10～20)%,而细胞工厂中的细胞几乎没有脱落;收获至第9次时,细胞工厂中的细胞脱落率才达20%;每次收获的病毒液病毒滴度和抗原含量均较高,且稳定在一定范围。浓缩纯化后,各项检定指标均合格。结论使用细胞工厂培养沙鼠肾原代细胞制备HFRS疫苗不仅提高了收率,而且减少了培养空间,降低了污染,可替代转瓶大规模生产HFRS疫苗。
Objective To prepare inactivated hemorrhagic feverwith renalsyndrome（HFRS）vaccine by primary kidney cells of gerbil mice cultured on Cell-Stack apparatus（also called cell factory）instead of typical roller culture so as to increase the yield and decrease the use of gerbil mice.Methods Primary kidney cells of gerbil mice were cultured on Cell-Stack apparatus,using those cultured in 15 L rolling bottle as control.The growths of cells,as well as titers and antigen contents of harvested virus cultured by the two methods were compared.The harvested virus liquid was concentrated by ultrafiltration,then purified by Sepharose-4FF chromatography and subjected to control tests.Results At the same stage of culture,the quantity of cells on Cell-Stack apparatus was equal to that in rolling bottle,while the quantity of primary kidney cells of gerbil mice inoculated required by the former was only 1 /3 of that by the latter.About（10 ～ 20）% of cells inoculated with virus in roller culture were exfoliated after harvest for 5 times,while those cultured on Cell-Stack apparatus showed little exfoliation,only 20% of which were exfoliated after harvest for 9 times.The titer and antigen content of each virus harvest of culture on Cell-Stack apparatus were higher than those of roller culture,which were stable within a certain range.The virus liquid after concentration and purification was qualified in control tests.Conclusion The yield of vaccine prepared by culture of primary kidney cells of gerbil mice on Cell-Stack apparatus increased,while the cultural space and contamination decreased,indicating that Cell-Stack apparatus might be used for large-scale production of inactivated HFRS vaccine instead of roller culture.
出处
《中国生物制品学杂志》
CAS
CSCD
2010年第6期613-614,618,共3页
Chinese Journal of Biologicals
关键词
肾综合征出血热
细胞工厂
沙鼠亚科
肾
细胞
Hemorrhagic fever with renal syndrome（HFRS）
Cell-Stack apparatus（Cell factory）
Gerbillinae
Kideny
Cells
