新生牛血清的质量控制对SPF鸡胚细胞和人二倍体(2BS)细胞生长的影响  

Effect of quality control of newborn bovine serum on growths of SPF chick embryo cells and human diploid (2BS) cells

作　　者：付永其 金珏艳 张颖欣 刘辉 陆晨 宋艳梅 杨琦 陆梦溪 王为 FU Yong-qi;JIN Jue-yan;ZHANG Ying-xin;LIU Hui;LU Chen;SONG Yan-mei;YANG Qi;LU Meng-xi;WANG Wei(Beijing Bei Shengyan Biological Products Co.,Ltd.,Beijing 100176,China)

机构地区：[1]北京北生研生物制品有限公司,北京100176 [2]浙江天杭生物科技股份有限公司,浙江湖州330521

出　　处：《中国生物制品学杂志》2018年第11期1182-1187,1191,共7页Chinese Journal of Biologicals

基　　金：疫苗综合性技术研究开发大平台建设(2013ZX09402302)

摘　　要：目的考察新生牛血清质量控制对SPF鸡胚细胞和人二倍体(2BS)细胞生长的影响。方法选择来源可追溯性、工艺质量稳定,且检定合格的新生牛血清样品配制细胞生长液,用于SPF鸡胚细胞和2BS细胞的培养。待细胞生长成致密成纤维单层细胞,记录细胞成单层的时间,用胰酶消化细胞,通过全自动细胞计数仪对细胞悬液进行测定,观察总细胞数、活细胞数,计算细胞存活率。根据活细胞数计算MOI,分别用腮腺炎及风疹病毒接种SPF鸡胚细胞和2BS细胞,检测收获病毒液的滴度。结果SPF鸡胚细胞总细胞数不低于3.5×10~6个/mL,活细胞数不低于2.1×10~6个/mL,细胞存活率不低于60%;2BS细胞总细胞数不低于7.0×10~5个/mL,活细胞数不低于4.2×10~5个/mL,细胞存活率不低于60%。腮腺炎病毒原液滴度在6.1～6.6 lgCCID_(50)/mL之间,平均值为6.28 lgCCID_(50)/mL,SD为0.11,生产过程加工能力指数(process capability index,Cpk)为1.51,批间质量趋于一致;风疹病毒原液滴度均在5.5～6.2 lgCCID_(50)/mL之间,平均值为5.8 lgCCID_(50)/mL,SD为0.17,Cpk值为1.40,批间质量趋于一致。结论新生牛血清的质量良好,用其培养SPF鸡胚细胞和2BS细胞的活细胞比率可控,收获的病毒液滴度较均一,差异性较小。Objective To investigate the effect of quality control of newborn bovine serum on growths of SPF chicken embryo cells and human diploid（2BS）cells.Methods Cell growth mixture was prepared with qualified newborn bovine serum with traceable source and stable process quality,and used for the cultivation of SPF chicken embryo cells and 2BS cells.After formation of a dense fibroblast monolayer,the time for formation was recorded,and the cells were digested with trypsin.The total and viable cells in cell suspension were measured by using automatic counter,and the cell survival rate was calculated.According to the viable cell count,MOIs were calculated,at which mumps and rubella viruses were inoculated into SPF chicken embryo cells and 2BS cells respectively.The harvested virus liquids were determined for titer.Results The total count,viable count and survival rate of SPF chicken embryo cells were not less than 3.5×10^（6 ）cells/mL,not less than 2.1×10^6cells/mL and not less than 60%,while those of 2BS cells were not less than 7.0×10^（5 ）cells/mL,not less than 4.2×10^5cells/mL and not less than 60%,respectively.The mean titer of bulk of mumps virus was6.28 lgCCID_（50）/mL,with a SD of 0.11 within a range of 6.1～6.6 lgCCID_（50）/mL,while the process capability index（Cpk）was 1.51,indicating consistent qualities between various batches.However,the mean titer of bulk of rubella virus was 5.8 lgCCID_（50）/mL,which was within the range of 5.5～6.2 lgCCID_（50）/mL,with a SD of 0.17 and a Cpk of 1.40,indicating consistent qualities between various batches.Conclusion The newborn bovine serum showed good quality,with which the survival rates of cultured SPF chicken embryo cells and 2BS cells were controllable.The titers of harvested viruses showed no significant difference.

关 键 词：新生牛血清 质量控制 SPF鸡胚细胞 人二倍体(2BS)细胞 细胞存活率 病毒滴度 

分 类 号：R392-33[医药卫生—免疫学]