牛血清在百日咳毒素CHO细胞簇聚试验中的影响  

Effect from fetal calf serum on Chinese hamster ovary cell clustering assay in test of pertussis toxin

作　　者：刘晓凤[1] 邓杰[2] 秦敏[1] 

机构地区：[1]成都生物制品研究所有限责任公司细菌性疫苗一室,四川成都610023 [2]成都生物制品研究所有限责任公司,四川成都610023

出　　处：《微生物学免疫学进展》2016年第2期50-54,共5页Progress In Microbiology and Immunology

摘　　要：目的观察不同胎牛血清对百日咳毒素在中华仓鼠卵巢细胞（Chinese hamster ovary cell,CHO）细胞簇聚试验中的影响。方法分别用两个厂家共6批次的牛血清培养CHO细胞,连续传3代后进行细胞簇聚试验,观察添加PT阳性对照、纯化PT、脱毒PT后其细胞的簇聚效果。结果 1、2号牛血清培养的CHO细胞生长缓慢,3-6号牛血清培养的CHO细胞生长正常。质量浓度16 ng/m L的PT阳性对照和纯化PT均未引起1-2号CHO细胞簇聚;3-6号牛血清培养的CHO细胞PT阳性对照判定终点分别为2、1、16、4 ng/m L,其纯化PT判定终点分别为1、0.5、8、2 ng/m L;脱毒PT质量浓度为40μg/m L时,1、2、5号牛血清培养的CHO不簇聚,而3、4、6号牛血清培养的CHO细胞脱毒PT判定终点分别为10、5、20μg/m L。结论 6种牛血清培养的CHO细胞簇聚程度存在差异,其中以4号牛血清培养的CHO细胞对PT阳性对照、纯化PT和脱毒PT最为敏感。需筛选对簇聚试验敏感度高的牛血清用于百日咳毒素CHO细胞簇集试验。Objective To compare the effect from different lots of bovine serum on Chinese hamster ovary cell（ CHO） clustering assay for pertussis toxin. Methods CHO cells were culutured with 6 lots of fetal calf serum from two manufacturers,after processed 3 passages,the cells were used for the clustering assay,and observation was carried out in clusterin result for added PT positive control,purified PT,detoxified PT. Results The cells growed in a slow state by using of No. 1 and No. 2 serum,and cells growed in a normal state by using of No. 3- No. 6. As the concentration of PT positive control and purified PT was used in 16 ng / m L,CHO cells did not occur clustering by using No1 and No2 serum. The end point for PT positive control were 2,1,16,4 ng / m L for No3- 6 serum,respectively,and 1,0. 5,8,2 ng / m L for thier purified counterpart. When the detoxified PT concentration was 40 μg / m L CHO cells did not occur clustering by using of No1,2,5 serum,respectively,but the clustering occurred with end point 10,5,20 μg / m L with No. 3,4,6 serum,respectively. Conclusion There were some differences in CHO cell clustering by using of 6 lots of fetal calf serum,it was the most sensitive to PT positive control,puriefied PT and detoxified PT with No. 4 serum. Search for the most sensitive fetal calf serum in culture of CHO cell should be performed in beforehand for the related clustering assay.

关 键 词：胎牛血清 中华仓鼠卵巢细胞 簇聚试验 

分 类 号：R392-33[医药卫生—免疫学]