经阿魏酸钠定向诱导的骨髓间充质干细胞在脑缺血大鼠体内移植研究

目的探讨经阿魏酸钠定向诱导的人骨髓间充质干细胞(mesenchymal stem cells,MSC)体内移植后在局灶性脑缺血大鼠脑内的存活及分布状况。方法分离、培养人MSC,经阿魏酸钠定向诱导后用于移植。复制大鼠局灶性大脑中动脉栓塞模型,实验组经股静脉注入诱导后CM-DiI标记的MSC,对照组注入等体积的生理盐水。2周后取脑组织行冰冻切片,荧光显微镜下观察移植细胞在脑组织中的存活及分布状况,同时检测神经元标志物NSE的表达。结果 MSC经阿魏酸钠诱导24h后,细胞逐渐圆缩并伸出突触,呈现神经细胞样形态,CD29表达阳性,CD34表达阴性。实验组大鼠脑内可见CM-DiI标记的MSC,主要分布在缺血侧损伤灶;同时可检测到细胞表面表达神经元标志物NSE。结论 MSC经阿魏酸钠诱导后向神经细胞分化,诱导分化的细胞能在脑缺血大鼠脑内存活,且主要分布在缺血侧损伤区,并仍保留已分化神经细胞的特性。

Transplantation of bone marrow mesenchymal stem cells induced by sodium ferulate into rats with cerebral ischemia

Objective To investigate the survival and distribution after implantation of human bone marrow mesenchymal stem cells （BM-MSCs） induced by sodium femlate （SF） in the brain in middle cerebral artery occlusion （MCAO） rat. Methods Human BM-MSCs were isolated, cultured and induced by SF into neuron-like cells, which was confirmed by fluorescent immunocytochemistry. Then the cells labeled with fluorescent dye CM-DiI were transplanted into MCAO model of SD rats through femoral vein in the experimental group and equal volume of physiological saline was injected in the control group. Frozen sections of the brain samples were prepared 2 weeks after transplantation. The survival and distribution of transplanted cells were observed under fluorescence microscope and the expression of neuron-specific enolase （NSE） was detected by immunofluorescence staining. Results Twenty-four hours after induction, the induced cells gradually became round and shrank with synapse formation and displayed a neuron-like morphology. Immunohistochemistry was positive for CD29 and negative for CD34. The CM-DiI-labeled BM-MSCs and NSE-positive cells were observed mainly in the ischemia injury regions of rat brain. Conclusions BM-MSCs can be induced by SF into neurons, which survive, mainly locate in the isehemia injury regions of MCAO rat brain and can remain the characteristics of differentiated neurons.