绿色荧光蛋白腺病毒表达载体在肌源性干细胞移植修复脊髓损伤实验中的应用

目的:观察绿色荧光蛋白腺病毒表达载体在肌源性干细胞移植修复脊髓损伤实验中的应用。方法:40只成年SD大鼠随机分为移植组(n=20)和对照组(n=20),均进行脊髓半切损伤。伤后9d,移植组于伤处移植体外转染GFP基因的大鼠MDSCs,而对照组仅注射等量PBS,于移植后1、2、3、4周用BBB评分方法测大鼠的运动功能,同时进行损伤脊髓取材、快速冰冻切片进行荧光显微镜观察。结果:移植后1周两组动物运动功能恢复无明显差异;移植后2、3、4周与对照组比较,移植组显著提高运动功能;荧光显微镜观察经绿色荧光蛋白基因标记的肌源性干细胞在损伤脊髓组织局部生长良好,并且有沿着脊髓神经束向头尾两侧迁移的趋势。结论:携带绿色荧光蛋白基因的腺病毒表达载体的细胞标记方法是安全可靠、简便有效的生物示踪标记方法,可以用于标记细胞移植实验研究。

Application of green fluorescent protein adenovirus expression vector on labeling muscle-derived stem cells' and Transplantating on spinal cord injury of rats

Objective：To study the green fluorescent protein（GFP） expression vector’s trace labeling effect to muscle-derived stem cells’ transplantating on Spinal Cord Injury of Rats.Methods：Forty adult Sprague-Dawley rats were randomly divided into transplantation group（n=20） and control group（n=20）,and spinal cord hemisection was performed in all rats.Nine days post-injury,MDSCs with GFP gene were transplanted into the injured spinal cord in transplantation group while control group was treated with injection of phosphate buffer saline（PBS）.Then the rats’ behaviors were assessed by BBB score 1w,2w,3w,and 4w after transplantation respectively.At the same time the injuried spinal cords were made into frozen sectiones and were observed through fluorescence microscope.Results：As compared with control group,MDSCs treatment significantly improved the locomotor recovery of hSCI in rats 2w,3w,4w post-transplantation respectively.Observed under fluorescence microscope Muscle derived stem cells marked with GFP grow well at the injuried myeloid tissue and transference following spinal nerves tract.Conclusion：The way that sign the cells with adenovirus expression vector that taking along green fluorescent protein is a safety convenient effective biologic trace labeling method,the signed cells can be used in the cell transplantation therapy experimentation.