聚乙烯亚胺介导骨形态发生蛋白2基因转染骨髓间充质干细胞的研究  

Polyethylenimine-mediated transfection of bone marrow mesenchymal stem cells

作　　者：方志辉[1] 谭金海[2] 简超[2] 

机构地区：[1]湖北医药学院附属东风医院创伤外科,湖北十堰442000 [2]武汉大学中南医院骨外科

出　　处：《中华实验外科杂志》2012年第11期2268-2270,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省自然科学基金资助项目（2009CDA058）

摘　　要：目的观察聚乙烯亚胺（PEI）转染骨髓间充质干细胞（BMSCs）的效率，从而优化可用于转染实验的条件。方法利用细胞计数试剂盒（CCK-8）比色法检测0．2％-2．0％PEI的细胞毒性，利用荧光显微镜和流式细胞仪测定NIP3．97-37．75的转染效率、培养液中的氯喹（0-100μmol／L）、10％白蛋白、5％血清及0-20mmol／LMg^2＋对PEI转染BMSCs效率的影响。结果当完全培养基内的PEI浓度低于0．8％，BMSCs存活率为70％；N／P15时PEI转染BMSCs的效率为14．4％；溶酶体抑制剂氯喹可增加PEI的转染效率，培养液中的10％白蛋白、5％血清及Mg^2＋可降低PEI转染效率。结论PEI是一种有效的BMSCs非病毒转染剂。Objective To study transfection efficiency of polyethylenimine （PEI） into bone mar- row mesenchymal stem cells （BMSCs） in order to optimize the conditions that can be used for transfection experiments. Methods Cell counting kit-8 （CCK-8） assay was used to test the cytotoxicity of 0. 2%- 2.0% PEI. Flow cytometry and fluorescence microscope were employed to determine the transfection effi- ciency by N/P 3.97-37. 75 and test the effects of ehloroquine （0-100 p^mol/L） , 10% albumin, 5% serum and 0-20 mmol/L Mg2＋ in the culture on the transfection efficiency of BMSCs. Results When PEI in the complete culture medium was below 0. 8%, the survival rate of BMSCs was 70%. After optimizing the N/P charge ratio, the PEI/DNA microcapsule delivered plasmid into BMSCs and obtained optimum transfection efficiency N/P 15. Chloroquine enhanced the transfection efficiency of PEI. 10% albumin, 5% serum and Mg2＋ in the culture decreased the transfection efficiency. Conclusion PEI is a kind of effective non-viral vectors for BMSCs.

关 键 词：非病毒基因载体 转染 聚乙烯亚胺 骨髓间充质干细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]