阳离子脂质体介导的基因转染大鼠BM-EPC的研究  

Transfection of Foreign DNA into Bone Marrow-Endothelial Progenitorcells (BM-EPCs) Mediated by Cationic Liposome

作　　者：夏爽[1] 佘强[1] 张玉[1] 邓松柏[1] 汪洋[2] 

机构地区：[1]重庆医科大学第二附属医院心血管内科,重庆400010 [2]重庆医科大学公共卫生学院卫生毒理学教研室,重庆400010

出　　处：《生物技术》2009年第3期19-21,共3页Biotechnology

基　　金：重庆市教委资助课题(渝教科[2004]12号)

摘　　要：目的:探讨阳离子脂质体法对体外分离培养的Sprague-Dawley(SD)大鼠骨髓来源-血管内皮祖细胞(Bone Marrow-En-dothelial progenitorcells,BM-EPCs)进行基因转染时脂质体和DNA质粒安全有效的剂量组合。方法:体外分离、培养SD大鼠BM-EPCs;免疫荧光技术测定CD34、CD133、Dil-acLDL,对细胞进行鉴定;按照正交设计,用不同水平的脂质体和质粒pEGFP转染细胞;荧光显微镜下计数阳性转染细胞,计算转染率。结果:SD大鼠BM-EPCs细胞表面抗原CD133、CD34呈阳性表达并具有吞噬Dil-acLDL的功能,形态学上两种细胞亚型-早期及晚期外生EPCs共存于其中。Lipofectamine 2000和pEFGP不同剂量组合均可转染大鼠BM-EPCs,其中脂质体5μl/质粒8μg时的转染效率高于二者其它剂量组合(P<0.05)。结论:优化条件后的阳离子脂质体Lipofectamine 2000可安全、有效转染体外分离培养的SD大鼠BM-EPCs。Objectlve：To explore the most optimized eombination of Lipofectamine 2006 and plasmid pEGFP when transfecting foreign DNA into Endothelial progenitorcells（EPCs） originated from rat＇s bone marrow. Method：EPCs were isolated from SD rat bone marrow,cultured in vitro as described in literature. The expression of CD133, CD34 and Dil - acLDL was conducted to identify EPCs, throughimmunofluorescent staining. Different levels of LipofectAMINE2000 and plasid pEGFP was transfected into EPCs according to orthogonal design, and cells with positively expressed GFP were counted under fluorescent microscope. Result： Cell markers CD34, CD133 and Dil - acLDL were all expressed in EPCs originated from rat＇s bone marrow; besides, it was found that early- and late- outgrowth EPCs coexisted when morphology was concerned. EPCs could be transfected by different dose combination of Lipofeetamine 2000 and, while the highest level of transfection was induced by 5μl of Lipofectamine 2000 and 8μg of pEGFP（p 〈 0.05）. Conclusion： Lipofeet AMINE 2000 is a safe and efficient way of EPCs transfection at the optimized dose.

关 键 词：内皮祖细胞 转染 阳离子脂质体 基因治疗 

分 类 号：R541.4[医药卫生—心血管疾病]