神经干细胞的培养鉴定及分化

发布时间：2018-07-03 00:03

  本文选题：神经干细胞 + 细胞　； 参考：《中国组织工程研究》2017年17期

【摘要】：背景:神经干细胞在神经损伤修复和退行性疾病中有广泛的应用前景,体外培养鉴定及促神经元诱导分化是后续研究的基础。目的:采用悬浮神经球培养法分离培养神经干细胞并对其鉴定,了解生物学特性。方法:采用悬浮神经球培养法从C57BL/6胎鼠大脑半球分离培养神经干细胞,观察形态特征及超微结构,CCK-8法绘制生长曲线,流式细胞仪检测细胞周期,免疫荧光法检测特异性标志蛋白Nestin的表达;用体积分数为1%和10%的血清诱导分化后,免疫荧光法检测GFAP、βⅢ-tubulin和MBP的表达。结果与结论:(1)体外培养得到悬浮生长的神经球,生长曲线和细胞周期表明细胞增殖力强;(2)透射电镜观察到神经干细胞核浆比高,呈未分化状态;(3)Nestin免疫荧光阳性;(4)不同体积分数的血清诱导后均可分化为星形胶质细胞、神经元和少突胶质细胞,体积分数为1%血清能诱导分化得到更多的神经元细胞;(5)结果表明,采用悬浮神经球培养法成功分离培养得到了神经干细胞,低体积分数血清有利于神经干细胞向神经元分化。
[Abstract]:Background: neural stem cells (NSCs) are widely used in nerve injury repair and degenerative diseases. In vitro culture and identification of neural stem cells and induction of neuronal differentiation are the basis of further research. Objective: to isolate and identify neural stem cells (NSCs) by suspension culture. Methods: neural stem cells were isolated and cultured from the cerebral hemisphere of C57BL / 6 fetal mice by suspension ball culture method. The morphology and ultrastructure of neural stem cells were observed and the growth curve was drawn by CCK-8 method. The cell cycle was measured by flow cytometry. The expression of nestin, a specific marker protein, was detected by immunofluorescence, and the expression of GFAP, 尾 鈪，

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