成人脑神经干细胞体内外增殖、分化和迁移研究  

Study on the proliferation,differentiation and migration of neural stem cells derived from adult brain in vitro and in vivo

作　　者：马献昆[1,2] 李建华[1] 宋丽惠[1] 郭欣茹[2] 王淑为[2] 刘爽[2] 

机构地区：[1]新疆生产建设兵团医院神经外科,新疆乌鲁木齐830002 [2]海军总医院神经外科,北京100048

出　　处：《中华神经外科疾病研究杂志》2013年第5期421-425,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(81172384)

摘　　要：目的探索从成人脑组织获取的神经干细胞(成人-hNSCs)在体外的增殖能力、分化特性、以及在裸鼠颅内的存活、迁移及分化情况。方法分别留取癫痫患者手术切除的颞叶脑组织和10 w左右人类自然流产胎儿纹状体组织,体外分离成单细胞悬液,无血清培养基培养、传代并诱导分化。软琼脂糖集落形成实验检测NSCs的增殖能力。免疫荧光法检测NSCs标志物神经上皮巢蛋白(Nestin)和诱导分化后神经元标志物β-tubllin以及神经胶质细胞标志物胶质纤维酸性蛋白(GFAP)的表达;利用动物立体定向仪将体外悬浮培养2 w的人NSCs移植入裸鼠颅内,检测NSCs在裸鼠脑组织局部的存活、迁移和分化状况。结果成人-hNSCs集落形成能力较胚胎脑组织来源的NSCs(胎儿-hNSCs)明显减弱,免疫荧光染色显示分离的NSCs呈Nestin阳性,诱导分化后可见(β-tubllin和GFAP阳性的神经细胞,其中80%的细胞为GFAP阳性的星形胶质细胞,20%左右为β-tubllin阳性细胞。分别将成人-hNSCs和胎儿-hNSCs移植入裸鼠纹状体,1个月后,冰冻切片,荧光显微镜观察到来源于成人脑组织来源的NSCs仅见沿针道的近距离迁移,免疫荧光染色在成人-hNSCs移植裸鼠颅内只检测到GFAP阳性的星形胶质细胞。而胎儿-hNSCs可穿过针道,沿大脑廉向脑实质广泛迁移,免疫荧光染色能检测到GFAP和少量(β-tubllin阳性细胞。结论成人脑组织和胚胎纹状体组织中均能成功分离到神经前体细胞,而与胎儿-hNSCs相比,成人-hNSCs体外增殖能力、多向分化潜能和体内迁移能力都明显减弱。Objective To investigate the proliferation and differentiation of human adult neural stem cells （adult-hNSCs） in vitro and in vivo. Methods Central nervous system-derived human neural stem cells （CNS- hNSCs） were obtained from adult human brain tissues and human aborted fetus brain （ 10 w） （fetus-hNSCs）, and were suspension cultured, passaged and induced to differentiate in vitro. NSCs marker-Nestin, neuron marker β- tublin and glial cell marker-glial fibrilary acidic protein （GFAP） were detected by method. Furthermore, the properties of NSCs in vivo was studied in nude mice rr^el. Results Compared with fetus-hNSCs, the ability of colony formation of the adult-NSCs was decreased significantly. Using the iinmunofluorescence method, nestin could be identified in adult-hNSCs and the differentiated cells expressed β-tublin and GFAP. After being transplanted into the corpus striatum of nude mice, adult-hNSCs could be detected to survive at least for 6 w, and differentiate into astrocytes. Conclusion CNS-hNSCs can be derived from adult brain and fetus brain. They could be cultured, passaged and induced to differentiate in vitro successfully. Compared with fetus-hNSCs, adult-hNSCs have limited abilities in proliferation, differentiation and migration in vivo.

关 键 词：神经干细胞 成人脑组织 增殖 分化 迁移 

分 类 号：R617[医药卫生—外科学]