Amaxa Nucleofector^(TM)核转染仪转染白血病细胞株L1210  

Amaxa Nucleofector^(TM) Nuclear Transfection Apparatus Transfers L1210 Cell Line

作　　者：杨阿碰[1] 马洁娴[1] 秦文明[2] 谢彦晖[1] 金由辛[2] 

机构地区：[1]复旦大学华山医院血液科,上海200040 [2]中国科学院上海生命科学研究院生物化学与细胞研究所,上海200031

出　　处：《中国实验血液学杂志》2010年第6期1604-1608,共5页Journal of Experimental Hematology

摘　　要：慢性淋巴细胞白血病细胞株L1210是广泛运用于白血病发病机制以及化疗药物研究的细胞模型,但该细胞株与其它悬浮细胞株一样,存在着转染效率低的难题。本研究旨在探讨如何提高L1210细胞株的转染效率。应用报告基因pEGFP并通过Amaxa NucleofectorTM核转染仪的几种转染预设模式转染悬浮L1210细胞,与脂质体Lipofectamine 2000转染进行对比,在荧光倒置显微镜下观察、使用流式细胞术检测转染效率,用台盼蓝排斥实验比较存活率。结果表明:使用核转染仪进行转染在L1210细胞中获得了很高的转染效率,且在一样细胞密度(2×106/ml)和质粒量(10μg)条件下,在24小时后进行比较,核转染仪预设模式A-20转染效率达(61.6%),明显高于其它模式(S-18、T-20),存活率可达50.5%;脂质体转染24小时细胞存活率为88%,但转染效率极低(<1%)。结论:核转染仪转染L1210的效率明显高于脂质体转染方案,其转染效率最高可达61.6%,存活率可达50.5%,此结果可满足科研工作需求。Mouse L1210 leukemia cell line is widely used as a model in the study of tumorigenesis, as well as the efficacy of chemotherapeutic drugs; however, like other suspension cell lines, the mouse L1210 cell line has lowest transfection efficiency, that many barriers exist to study about the structure, function, as well as metabolism in leukemia cells. This study was aimed to obtain higher transfection efficiency of L1210 cell line to facilitate scientific research. The transfection efficiencies of nucleofector and liposome in L1210 leukemia cells were detected by converted fluorescence microscopy and flow cytometry using EGFP （enhance green fluorescent protein）; cell viability was observed by trypan blue exclusion test. The results showed that the transfection efficiency of nucleofector primarily through reporter gene pEGFP by Amaxa NucleofectorTM nuclear transfer apparatus was significantly higher than lipofectamine 2000 transfection, furthermore, in the same cell density （2×106/ml） and plasmid content （10 μg）, the transfection efficiency of nuclear transfer apparaturs default mode A-20 was higher than that of other modes （S-18, T-20）. Its survival rate was up to 50.5% after 24 hours. Cell viability of liposome transfection reached to 88% after 24 hours, but the transfection efficiency was lower （1%）. It is concluded that the nuclear transfer apparatus A-20 transfected L1210 can reach higher transfection efficiency up to 61.6%, which is significantly higher than that of lipofectamine transfection. The survival rate is up to 50.5% well meeting the needs of scientific research. Higher transfection efficiency is helpful for in-depth research about the morphology, functions and pathogenesis in leukemia model L1210, and provides more searching space for the treatment of leukemia diseases.

关 键 词：L1210 EGFP 核转染 脂质体转染 

分 类 号：R733.7[医药卫生—肿瘤] Q789[医药卫生—临床医学]