磁性纳米颗粒与脂质体介导REST-siRNA转染hADSCs的比较  

Comparison of magnetic iron oxide nanoparticles and lipofectamine^(TM) 2000 connect REST-siRNA transfect human adipose mesenchymal stem cells

作　　者：王瑞[1] 庞希宁[1] 施萍[2] 

机构地区：[1]中国医科大学卫生部细胞生物学重点实验室干细胞与再生医学研究室,辽宁沈阳110001 [2]中国医科大学附属第一医院全科医学教研室,辽宁沈阳110001

出　　处：《基础医学与临床》2013年第12期1549-1553,共5页Basic and Clinical Medicine

基　　金：国家重点基础研究发展规划(2012CB518103);辽宁省科学技术计划(2012225080);沈阳市科学技术计划(F11-262-9-01)

摘　　要：目的对聚乙烯亚胺包被的磁纳米颗粒(PEI-SPIO)与脂质体(lipofectamineTM2000)作为基因载体转染人脂肪间充质干细胞(hADSCs)的可行性及转染后细胞的增殖、转染效率及基因瞬时干扰效率进行比较分析。方法分别将PEI-SPIO和lipofectamineTM2000作为基因载体连接REST-siRNA体外转染hADSCs,用MTT法检测hADSCs增殖和功能;用荧光显微镜观察转染效率;分别用Real-time PCR和Western blot检测REST的表达。结果用磁性纳米颗粒和脂质体作为基因载体转染细胞后,细胞成活率分别为99%和79%(P<0.05);磁纳米颗粒和脂质体的转染效率无差异,分别为75%和71%;磁纳米颗粒和脂质体作为基因载体时RNA表达量分别为(0.46±0.04)和(0.53±0.05),蛋白水平表达量分别为(0.501±0.006)和(0.523±0.004)。结论磁性纳米颗粒因其具有低细胞毒性可代替脂质体作为转染干细胞的基因载体。Objective Comparison of polyethyleneimine package magnetic nanoparticles and lipofectamineTM 2000 in transfection of human adipose mesenchymal stem cells.And the feasibility, instantaneous interference efficiency and cell proliferation state, the transfection efficiency were compared. Methods The PEI-SPIO and lipofectamineTM 2000 connect REST-siRNA in vitro and transfect into human adipose mesenchymal stem cells, MTT method for determin the effects of human adipose mesenchymal stem cell proliferation and function. transfection efficiency was detected by fluorescence microscope.And REST changes were determined with Real-time PCR and Western-Blot. Results The survival rate of the PEI-SPIO method was 99% and the lipofectamineTM 2000 method was 79%（P〈0.05）; the transfection efficiency of PEI-SPIO was no difference, 75%和71% respectively, PEI-SPIO and lipofectamineTM 2000 as gene carrier at RNA level was （0.46±0.04） and （0.53±0.05） respectively,and at protein level was （0.501±0.006） and （0.523±0.004） respectively. Conclusion Magnetic nanoparticles because of its low toxicity can replace the lipofectamineTM 2000 transfect stem cells as a gene vector.

关 键 词：磁性纳米颗粒 聚乙烯亚胺 脂质体 脂肪间充质干细胞 转染 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]