不同来源成体神经干细胞神经生物学特性比较的实验研究  

Comparison of biological characteristics among adult neural stem cells derived from different origins in vitro

作　　者：蔡颖谦[1] 唐艳平[1] 张洪钿[1] 姜晓丹[1] 徐如祥[1,2] 

机构地区：[1]南方医科大学珠江医院神经外科,广东神经外科研究所,广东脑功能修复与再生重点实验室,广州510282 [2]北京军区总医院神经外科,北京100700

出　　处：《中华神经医学杂志》2010年第5期446-450,455,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（30901546）;广东省自然科学基金（9451051501002508）;广东省科技计划项目（20098030801214）

摘　　要：目的 比较和评价来源于同一SD大鼠成体骨髓、脂肪和大脑的神经干细胞在体外自我更新、向神经元或神经胶质细胞分化的能力,对比三者体外分泌神经营养因子的能力,进一步评估和对比它们在移植修复中枢神经系统损伤的潜能.方法 选取来源于同一SD大鼠成体脑室下区、骨髓和脂肪3个部位的组织,体外分别诱导分化为脑室下区源性神经干细胞（SVZ-NSCs）、骨髓源性神经干细胞（BM-NSCs）和脂肪源性神经干细胞（AD-NSCs）,比较三者的自我更新和分化情况;免疫细胞化学方法 、Western blot检测巢蛋白（nestin）、β-微管蛋白（β-tubulin）、GFAP和半乳糖脑苷脂（GalC）4种细胞表面标记物的表达：ELISA法定量检测体外培养3种组织来源的神经干细胞的BDNF和NGF的分泌水平.结果 3种组织来源基质细胞形成的神经球从外形观察无明显差异;体外扩增率由高到低依次为SVZ-NSCs、BM-NSCs和AD-NSCs.ELISA检测结果 表明3组细胞都能检测出BDNF和NGF分泌,但BM-NSCs组和AD-NSCs组明显低于SVZ-NSCs组,差异均有统计学意义（P〈0.05）;AD-NSCs组BDNF和NGF的水平均稍高于BM-NSCs组. 结论 在增殖能力、向神经细胞终极分化的能力和分泌神经营养因子的能力等方面,BM-NSCs和AD-NSCs均稍逊于中枢神经组织来源的SVZ-NSCs,但由于其具有可来源于自体、无免疫排斥和伦理道德问题等优点,可超越SVZ-NSCs成为目前最有临床应用前景的移植用种子细胞.AD-NSCs来源更为丰富,取材更为简便,也更容易被患者接受,可能是更好的选择.Objective To evaluate and compare the adult neural stem cells （NSCs） derived from the subventricular zone （SVZ）, adipose tissue （AD） and bone marrow （BM） in SD rat in terms of their morphologies, their potential of neural differentiation and their ability to secrete neurotrophins （NTs）.Methods Tissues from the suventricular zone, adipose tissue and bone marrow in the same SD rat were chosen and induced in vitro into SVZ-NSCs, AD-NSCs and BM-NSCs, respectively. The abilities of proliferation and differentiation of these 3 NSCs were compared. Immunocytochemistry and Western blotting were employed to detect the expressions of surface markers of neurons and neuroglia, including nestin,βtubulin, galactocerebroside C （GalC） and glial fibrillary acidic protein （GFAP）. The secretions of BDNF and NGF were detected by ELIS A. Results No obvious differences of morphology between SVZ-NSCs and both BM-NSCs and AD-NSCs were found （P〉0.05）. The proliferation ability of SVZ-NSCs was stronger than that of BM-NSCs and AD-NSCs. The percentage of nestin-positive cells in the SVZ-NSCs was significantly higher than that in the BM-NSCs or AD-NSCs （P〈0.05）. No obvious differences in the expressions of βtubulin, GFAP, and GalC among the 3 groups were found （P〉0.05）.The secretions of BDNF and NGF in all the 3 groups could be noted; those in the SVZ-NSCs was significantly higher than those in the BM-NSCs and AD-NSCs （P〈0.05）; those in the AD-NSCs was slightly higher than those in the BM-NSCs. Conclusions SVZ-NSCs, AD-NSCs and BM-NSCs show similar morphological and phenotypic characteristics; however, SVZ-NSCs present more powerful proliferation, differentiation and secretion abilities than AD-NSCs and BM-NSCs. Considering about such problems as immuno-repulsion, ethic and the origins, AD-NSCs appear to be the better choice than BSVZ-NSCs and M-NSCs.

关 键 词：神经干细胞 细胞分化 神经再生 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]