培养胚胎神经干细胞移植入纹状体后的形态学观察

本研究的目的是 :将体外培养的小鼠胚胎大脑皮层和脊髓的神经干细胞经单细胞悬液微移植后观察其在大鼠纹状体的存活、迁移和分化的状况。实验在无血清条件下将这些细胞扩增、培养再经活细胞荧光染料 Di I标记后 ,采用微移植的方法 ,通过脑立体定位仪上用微玻璃针将干细胞分别植入成年大鼠双侧纹状体的对称部位。大鼠存活 8周后 ,经灌注固定、恒冷箱切片 ,在荧光显微镜下观察标记的移植细胞的迁移状况 ;用星形胶质细胞特异性抗体观察移植区 GFAP的表达 ,以显示移植细胞分化状况。结果表明 :来源于胚胎小鼠大脑皮层和脊髓的体外培养神经干细胞经微移植后 ,均可在成年大鼠脑内纹状体区域存活 ,移植的神经干细胞可向周围的脑实质内迁移 ,迁移细胞沿特定的纹状体结构分布。神经干细胞主要分化成星形胶质细胞。本实验结果提示 ,移植的神经干细胞可在脑实质内存活、迁移和分化。

A MORPHOLOGICAL STUDY OF NEURAL STEM CELL AFTER TRANSPLANTATION INTO THE ADULT RAT STRIATUM

The purpose of the present study was to investigate the survival, migration and differentiation of neural stem cells which were isolated and expanded in vitro from mouse cerebral cortex and spinal cord after being transplanted into adult rat with single-cell suspension by micro-transplantation method. The cultured neural stem cells in serum-free medium from embryo mouse cortex and spinal cord were expanded in vitro and labeled by DiI, and then labeled stem cells were transplanted into the adult rat brain striatum by micro-transplantation method as single-cell suspension with stereo-injection technique. After 8 w, the operated rats were fixed, and cryostat slice were obtained and the morphology of the transplanted stem cells and their migration were observed under fluorescence microscope. Astrocyte specific GFAP was detected by immunocytochemistry to show the differentiation fate of the transplanted stem cells. The results showed that neural stem cells from embryo mouse cortex and spinal cord which were expanded in vitro could survive, migrate into the adult rat striatum and were distributed along special striatum structure after being transplanted by micro-transplantation method, but most of them differentiated into glia cells. The results indicated that the transplanted neural stem cells can survive, migrate and differentiate in the rat striatum.