山羊神经干细胞的分离培养及体外诱导分化  

In vitro culture and differentiation of goat neural stem cells

作　　者：夏许可[1,2] 张伟 余杰锋 章莹[1] 

机构地区：[1]解放军广州军区广州总医院骨科医院,广东省广州市510010 [2]南方医科大学研究生院广东省,广州市510515 [3]广东冠昊生物科技股份有限公司,广东省广州市510530 [4]广州医科大学研究生院,广东省广州市510182

出　　处：《中国组织工程研究》2014年第23期3691-3695,共5页Chinese Journal of Tissue Engineering Research

基　　金：广东省科技计划项目(2010B031100017);广州市科技计划项目(2012J4100039)~~

摘　　要：背景:文献报道不同种类的神经调控因子及神经胶质细胞对神经干细胞分化成熟具有不同的作用,但大动物的神经干细胞培养报道则相对较少。目的:探究山羊神经干细胞的培养条件及体外诱导分化结果。方法:取幼羊脑组织分离神经干细胞后于神经干细胞全培养液中悬浮培养,后期行抗-巢蛋白一抗免疫组织化学染色鉴定,同时取坐骨神经分离培养许旺细胞;以血清诱导干细胞体外诱导分化后行抗-S100一抗、抗-胶质纤维酸性蛋白一抗、抗-MAP2一抗染色分别标记许旺细胞及神经胶质细胞,以未添加一抗做对照,计算图像灰度值与对照组进行统计学比较。结果与结论:成功分离培养抗-巢蛋白染色阳性神经球及抗-S100染色阳性许旺细胞,后期神经干细胞体外诱导分化得抗-胶质纤维酸性蛋白、抗-MAP2染色阳性细胞,其免疫组织化学染色灰度值显著高于对照组;分化产物抗-S100染色为阴性,其免疫组织化学染色灰度值显著低于对照组。结果表明山羊神经干细胞可经体外诱导分化为神经元及星形胶质细胞,未发现神经干细胞诱导分化为许旺细胞。BACKGROUND：Different types of nerve regulatory factors and glial cells have been reported to exert different roles in the differentiation and maturation of neural stem cells, but culturing neural stem cells in large animals is relatively rarely reported. OBJECTIVE：To explore the way for culturing goat neural stem cells and to detect the outcome after in vitro differentiation. METHODS：The neural stem cellwas separated and cultured from the newborn goat cerebral cortex and the anti-nestin immunocytochemical staining was performed for cellidentification. At the same time, anti-S100 active Schwann cells were gotten from the sciatic nerve. Then in vitro differentiation was preformed and the outcome was detected by the immunocytochemical stain of anti-glial fibril ary acidic protein, anti-microtubule-associated protein 2 and anti-S100. cells without primary antibodies served as controls. Gray values were calculated and compared. RESULTS AND CONCLUSION：The Schwann cells were cultured successful y, which were active to the anti-nestin immunocytochemical staining and anti-S100 staining. After differentiation, the products were active to anti-glial fibril ary acidic protein and anti-microtubule-associated protein 2 immunocytochemical stain, but Abstract BACKGROUND：Different types of nerve regulatory factors and glial cells have been reported to exert different roles in the differentiation and maturation of neural stem cells, but culturing neural stem cells in large animals is relatively rarely reported. OBJECTIVE：To explore the way for culturing goat neural stem cells and to detect the outcome after in vitro differentiation. METHODS：The neural stem cellwas separated and cultured from the newborn goat cerebral cortex and the anti-nestin immunocytochemical staining was performed for cellidentification. At the same time, anti-S100 active Schwann cells were gotten from the sciatic nerve. Then in vitro differentiation was preformed and the outcome was detected by the immunocytochemical stain of anti-glial fibril a

关 键 词：干细胞 培养 神经干细胞 干细胞培养 体外诱导分化 免疫组织化学 山羊 

分 类 号：R394.2[医药卫生—医学遗传学]