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目的 从胚胎大鼠脊髓神经管中分离神经上皮干细胞并诱导其向多巴胺能神经元方向分化。 方法 利用无血清悬浮培养、单细胞克隆技术分离神经上皮干细胞 ;采用 5 溴 2 脱氧尿苷 (BrdU)标记新生细胞 ,免疫细胞化学单标或双标染色技术 ,检测神经上皮干细胞蛋白 (nestin)和分化后特异性神经细胞抗原的表达 ;用纹状体组织提取液 ,诱导神经上皮干细胞向多巴胺能神经元方向分化。 结果 从胚胎大鼠脊髓神经管中分离的细胞可以连续传代 ,表达nestin ,它们分化后可以表达神经元、星形胶质细胞和少突胶质细胞的特异性抗原 ;与对照组 3%相比 ,纹状体组织提取液可以诱导这些细胞中的 12 %分化成为多巴胺能神经元。 结论 分离自脊髓神经管的细胞具有自我更新能力和多分化潜能 (multipotent) ,是增殖能力很强的神经干细胞 ;这些干细胞在一定的体外环境中能被诱导成为特定的神经元 ,提示其可以为神经移植提供材料
OBJECTIVE: To isolate neural epithelial stem cells from neural tube in embryonic rat spinal cord and induce its differentiation into dopaminergic neurons. METHODS: Neural stem cells were isolated by serum-free suspension culture and single cell cloning technique. BrdU-labeled neonatal cells, immunocytochemical single or double-stained staining were used to detect the expression of nestin and Differentiation of specific neuronal antigen expression; with striatum tissue extract, induced neural epithelial stem cells to dopaminergic neurons in the direction of differentiation. Results The cells isolated from the neural tube of embryonic rat spinal cord could be passaged continuously and expressed as nestin. After differentiation, they could express specific antigens of neurons, astrocytes and oligodendrocytes; compared with 3% of the control group , Striatal tissue extracts can induce 12% of these cells to differentiate into dopaminergic neurons. Conclusion The neural stem cells isolated from neural tube of spinal cord possess the ability of self-renewal and multipotent and are neural stem cells with strong proliferative ability. These stem cells can be induced into specific neurons in a certain environment in vitro, suggesting that they may be Nerve transplants provide the material