目的 通过腺病毒转染携绿色荧光蛋白(GFP)的脐带间充质干细胞(ucMSCs)移植大鼠脑缺血再灌注损伤模型,研究GFP作为示踪剂的可行性.方法 体外培养经腺病毒转染GFP的ucMSCs,建立大鼠脑缺血再灌注损伤模型,通过脑立体定位技术将1×106携GFP的ucMSCs移植入损伤脑组织,并于造模7 d后取脑组织进行冰冻切片,荧光显微镜下观察绿色荧光蛋白表达情况.结果 体外携GFP的ucMSCs形态呈成纤维状,融合时聚集成簇,呈放射状或漩涡状排列,与未转染ucMSCs形态无差异,荧光显微镜下可见GFP的阳性率达80%以上,且荧光扩散至整个细胞,可反映细胞完整形态.移植后大鼠未出现排斥反应,7 d后脑组织移植部位附近均可见GFP表达,荧光未见衰减.结论 GFP作为ucMSCs移植治疗脑损伤的有效示踪剂,可为细胞水平的各种研究及体内移植细胞的研究提供实验手段.%Objective To investigate the feasibility of green fluorescent protein (GFP) as a marker to trace the transplanted um-bilical cord mesenchymal stem cells (ucMSCs) in rats with cerebral ischemia-reperfusion .Methods ucMSCs were transfected by GFP-adenovirus .The rats were subjected to left middle cerebral artery occlusion using suture method .1 × 106 GFP-ucMSCs were transplanted with cerebral stereotaxic technique .Frozen sections of brain tissue were made at 7 d after cerebral ischemia .The ex-pression of GFP was observed by fluorescence microscope .Results In vitro ,the morphology of GFP in ucMSCs was fibrous ,and when fused ,the clusters were arranged in a radial or whirlpool shape ,The morphological feature of transfected ucMSCs was similar to that un-transfected ucMSCs .Under the fluorescence microscope ,the positive rate of GFP was more than 80% .In addition ,GFP could spread to the whole cells and show the completed form .No rejection was observed in the rats after transplantation ,and the GFP was found near the site of transplantation after 7 d ,and the fluorescence was not attenuated .Conclusion GFP is an effective tracer maker for ucMSCs transplantation in the treatment of ischemia-reperfusion ,which could provide experimental method for fur-ther study .
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