摘要
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CD 44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。
Objective：To observe the expression of bone marrow mesenchymal stem cells（BMSCs） in vitro, which is carried by plasmid and transfected by green fluorescent protein （GFP）. Methods： With plasmid virus as the cartier and PT 67 cells as the packaging cells, mediating GFP to trasfect the New Zealand rabbits. Cells were observed under the phase contrast microscope and fluorescence microscope. GFP expression efficiency was detected by flow cytometry instrument. Results：BMSCs were 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 transfeetion efficiency of GFP was above 30%. Conclusion： BMSCs can be easily obtained and proliferated, therefore, they may be applicated as seed-cells in osseous tissue engineering. However, GFP gene will not affect the activity and biological characteristics of BMSCs.
出处
《内蒙古医科大学学报》
2013年第2期96-101,共6页
Journal of Inner Mongolia Medical University
基金
内蒙古医科大学科技与工程项目(NY2011BW007)
内蒙古医科大学青年基金(NY 2004 QN09)
关键词
骨髓基质干细胞
GFP
基因转染
骨组织工程
bone marrow mesenchymal stem cells
green fluorescent protein
gene transfection
osseous tissue engineering
