绿色荧光蛋白体外转染兔骨髓基质干细胞greenfluorescent.pdfVIP

·96 · Journal Of InnerMongoliaMedical University摇 Apr.2013摇 Vol.35摇 No.2 绿色荧光蛋白体外转染兔骨髓基质干细胞 1,2 3 1 1 1 1 1 吕慧成 ,摇 李斯琴 ,摇 周荣兴 ,摇 李晓东 ,摇 郭摇 军 ,摇 贾海生 ,摇 吴一民 (1.重庆医科大学重庆市;摇 2. 内蒙古医科大学第二附属医院创伤二科;摇 3. 内蒙古人民医院超声科) 摇 摇 摘摇 要:目的:观察以质粒为载体,经脂质体介导的绿色荧光蛋白(green fluorescent protein,GFP)转染的骨髓 间充质干细胞体外表达情况。 方法:以质粒病毒为载体,PT 67 细胞为包装细胞,介导绿色荧光蛋白转染新西兰 大白兔,在相差显微镜和荧光显微镜下观察,流式细胞仪检测GFP 的表达效率。 结果:骨髓基质干细胞体外培 养后贴壁、集落生长,连续传代培养生长良好。 培养2 wk后骨髓基质干细胞表面CD44 抗原表达阳性。 介导绿 色荧光蛋白转染新西兰大白兔的骨髓基质干细胞,转染率可达30%左右。 结论:骨髓基质干细胞容易获取、来 源充足、可以大量扩增,可作为骨组织工程的种子细胞。 GFP基因的导入不会影响骨髓间充质干细胞的活性和 生物学特性。 关键词:骨髓基质干细胞;GFP;基因转染;骨组织工程 中图分类号:R392.4摇 摇 摇 摇 摇 文献标识码:A摇 摇 摇 摇 摇 文章编号:2095-512X(2013)02-0096-06 GREEN FLUORESCENT PROTEIN VITRO TRANSFECTION OF RABBIT BONE MARROW STROMAL STEM CELLS LU Hui-cheng,摇 LI Si-qin,摇 ZHOU Rong-xing,摇 et al. (Two Department of Trauma,The SecondAffiliated Hospital,InnerMongoliaMedical University,Hohhot010030 China) Abstract:Objective:To observetheexpressionof bonemarrow mesenchymal stemcells(BMSCs) in vitro,which is carried by plasmid and transfected by green fluorescent protein(GFP).Methods: With plasmid virus as the carrier and PT67 cellsasthe packaging cells,mediating GFP to trasfect the New Zealand rabbits. Cells were observed under the phase contrast microscope and fluorescence microscope.GFP expressionefficiencywasdetectedbyflow cytometry instrument.Results:BMSCswere colonial growth and even amplificated well in continuous passage culture.Additionally,CD44 on the surface of BMSCs positively expressed after 2 weeks cultured in vitro. Detected by flow cytometry instrument,the transfection efficiency of GFP was above 30%. Conclusion:BMSCs can be easily obtainedandproliferated,therefore,theymaybeapplicatedasseed-cellsinosseoustissueengineering.