无血清培养液在分泌鼠疫F1McAb杂交瘤细胞中的初步应用  

Preliminary Application of Serum-free Culture Medium in Hybridoma Secreting F1McAb of Yersinia pestis

作　　者：杜春红[1] 宋志忠[1] 唐雪[1] 钟佑宏[1] 周东升[2] 于国林[1] 

机构地区：[1]云南省地方病防治所,云南大理671000 [2]大理学院公共卫生学院,云南大理671000

出　　处：《地方病通报》2009年第1期7-9,13,共4页Endemic Diseases Bulletin

基　　金：云南省科技攻关项目(2007CA010)

摘　　要：目的应用ADCF无血清细胞培养液对分泌鼠疫F1McAb的杂交瘤细胞进行静止培养,初步了解该瘤株在ADCF中的生长情况,为今后采用无血清培养液体外培养法生产单克隆抗体提供依据。方法通过将杂交瘤细胞直接转入ADCF培养液培养、驯化培养和优化条件后培养,观察细胞在各种培养条件下的生长情况,进行活细胞计数和抗体效价检测。结果用ADCF培养鼠疫F1杂交瘤细胞,细胞直接转入不能较好的生长;通过驯化、在放过含血清培养液的原瓶中能较好的贴壁生长,在新瓶中不能稳定地贴壁生长和增殖;在经含血清的培养液优化后的培养瓶中,细胞能贴壁生长和增殖。结论ADCF培养液可用于分泌鼠疫F1McAb的杂交瘤细胞的培养。Objective To apply serum - free ADCF medium for the culture of hybridorna cell secreting F1 McAb of Yersinia pestis, so as to preliminarily understand the growing conditions of these strains, and further provide valid evidence for producing monoclonal antibody in external cell culture by using serum - free medium. Methods After directly applying ADCF medium for hybridoma cell culture, habituated culture and culture in optimizing conditions, the cell growth was observed under different conditions and counting live ceils and testing antibody titers were conducted as well. Results The growth of F1 hybridoma cell was poor when the cells were transferred to ADCF medium directly. It could grow adhering to the wall after habituated culture and in those bottles added the medium containing serum, whereas it failed to grow and multiply in new bottles. However, in those bottles optimized by medium containing serum, it began to grow and multiply. Conclusions ADCF medium can be used for the culture of the hybridoma cell secreting F1 McAb.

关 键 词：杂交瘤细胞 鼠疫菌 F1单克隆抗体 无血清培养 

分 类 号：R378.61[医药卫生—病原生物学]