NIH3T3-CD40L稳定细胞系构建及在EBV转化B淋巴细胞中的应用  

Construction of NIH3T3-CD40L Stable Cell Lines and Its Application in EBV Transformation of B Lymphocytes

作　　者：刘玺[1] 赵烨 许崇凤[1] 段子渊[1] Liu Xi;Zhao Ye;Xu Chongfeng;Duan Ziyuan(Genetic Resource Research Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China)

机构地区：[1]中国科学院遗传与发育生物学研究所遗传资源研究中心,北京100101

出　　处：《中国细胞生物学学报》2018年第5期768-774,共7页Chinese Journal of Cell Biology

摘　　要：通过脂质体法转染p CMV6-CD40L重组质粒和G418阳性克隆筛选,构建了稳定表达CD40L的NIH3T3细胞系,在RNA和蛋白质水平进行了功能鉴定,对其作为饲养细胞在体外B淋巴母细胞系(B lymphoblastoid cell lines,B-LCLs)培养和在EBV转化B细胞中的作用进行了实验验证。结果表明,NIH3T3-CD40L细胞系不仅可以提高体外低密度B-LCLs存活率,而且可以降低成功建立B-LCLs所需的细胞密度,促进其增殖。该细胞系的成功构建,进一步确证了NIH3T3-CD40L作为饲养细胞可以促进体外低密度B-LCLs的增殖及提高EBV转化B淋巴细胞的效率,优化了体外B细胞培养及EBV转化B淋巴细胞的流程,也为单克隆抗体的制备奠定了基础。Stably expressing CD40 L NIH3 T3 cell lines were constructed and its application as feeder cells to culture and establish B lymphoblastoid cell lines（B-LCLs） in vitro were evaluated. p CMV6-CD40 L was transfected into NIH3 T3 cell lines through lipofectamine method, followed NIH3 T3 that express CD40 L were screened by G418. Positive clones were confirmed at RNA and protein level, the NIH3 T3-CD40 L cell lines were used as feeder cells in culturing B-LCLs and in the process of EBV transformation. The results showed that NIH3 T3-CD40 L cell lines increased the survival rate and promote the proliferation of B-LCLs in low density and reduce the cell concentration required to establish B-LCLs. Therefore, NIH3 T3-CD40 L cell lines have been successfully constructed which would lay a foundation for further studies, such as monoclonal antibody production.

关 键 词：NIH3T3-CD40L 饲养细胞 B-LCLs 

分 类 号：R392[医药卫生—免疫学]