脂质体介导绿色荧光蛋白转染人脐血内皮祖细胞

发布时间：2018-03-09 19:50

本文选题：血管内皮祖细胞　切入点：绿色荧光蛋白　出处：《吉林大学》2009年硕士论文　论文类型：学位论文

【摘要】： 研究脂质体介导绿色荧光蛋白转染人脐血内皮祖细胞的方法的可行性,观察细胞荧光表达情况、转染效率及转染对细胞的影响。体外分离诱导培养人脐血内皮祖细胞(EPCs),用阳离子脂质体Lipofectamine2000介导携带有绿色荧光蛋白基因的质粒pEGFP-N1转入EPCs中,激光共聚焦显微镜观察荧光表达,全波长多功能酶标仪检测比较转染后24h、48h、72h、96h荧光强度,MTT法检测细胞数量变化,流式细胞术测定转染效率。绿色荧光蛋白在基因转染24h后已可见荧光表达,48h荧光表达明显增强,并且48h、72h、96h荧光强度呈递减趋势。转染后细胞数量减少。转染效率为21.6%±1.3%。脂质体可携带绿色荧光蛋白质粒转染内皮祖细胞,是标记内皮祖细胞的一种简便可行的方法。绿色荧光蛋白可在原代内皮祖细胞中表达。转染后细胞数量减少。
[Abstract]:To study the feasibility of liposome-mediated green fluorescent protein transfection into human umbilical cord blood endothelial progenitor cells, and observe the fluorescent expression of the cells. The efficiency of transfection and the effect of transfection on human umbilical cord blood endothelial progenitor cells were isolated and cultured in vitro. The plasmid pEGFP-N1 carrying green fluorescent protein gene was transfected into EPCs by cationic liposome Lipofectamine2000. The fluorescence expression was observed by confocal laser microscopy. After transfection, the number of cells was detected by MTT method and the transfection efficiency was determined by flow cytometry. The fluorescence expression of green fluorescent protein was obviously enhanced at 48 h after 24 h gene transfection. After transfection, the fluorescence intensity decreased, and the transfection efficiency was 21.6% 卤1.3. Liposomes could carry green fluorescent protein particles into endothelial progenitor cells. It is a simple and feasible method to label endothelial progenitor cells. Green fluorescent protein can be expressed in primary endothelial progenitor cells.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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