2种神经干细胞移植方法对宿主脑损伤和移植物维系的影响  

Influence of two different neural stem cells transplantation methods in brain injury and graft sustain in mice

作　　者：高谋[1] 徐如祥[2] 杨志军[2] 董勤[3] 张洪钿[2] 朱建伟[2] 刘宁[2] 邹明明[2] 籍新潮[2] 

机构地区：[1]第三军医大学大坪医院神经外科,重庆400038 [2]北京军区总医院附属八一脑科医院,北京100700 [3]首都医科大学附属复兴医院神经内科,北京100038

出　　处：《中华神经医学杂志》2015年第4期458-462,共5页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81271316）

摘　　要：目的比较2种神经干细胞（NSCs）颅内移植方法对宿主脑损伤和移植物维系的影响。方法体外原代培养NSCs并用5-溴脱氧尿嘧啶核苷（Brdu）标记，将NSCs单细胞悬液（5×10^5个，5μL）定向注射到42只健康成年C57BL／6小鼠两侧大脑运动皮层内，左侧颅骨采用磨钻钻孔，右侧采用微量进样器钻孔。移植后1d、2d、3d、7d、14d、21d和28d采用随机数字表法各选取6只小鼠处死，观察脑大体标本，HE染色观察脑移植区的损伤，免疫荧光双重标记染色Brdu／神经元核抗原（NeuN）、Brdu／神经胶质纤维酸性蛋白（GFAP）分析脑移植区NSCs、神经元和星形胶质细胞的分布情况。结果（1）与微量进样器钻孔侧相比，磨钻钻孔侧脑组织结构破坏较严重，移植后1～3d神经细胞坏死明显，并伴有大量红细胞外溢及炎症细胞浸润，7d时可见瘢痕组织形成，直到28d仍可见较多瘢痕组织聚集在移植区。（2）移植后1～3dBrdu标记的NSCs分布较为集中，7～21d可见NSCs明显迁移并分化为神经元和星形胶质细胞．左侧脑移植区可见大量星形胶质细胞．而神经元分布较少，右侧则相反；（3）与左侧比较，右侧脑移植区各时间点Brdu、NeuN阳性细胞百分率较高，GFAP阳性细胞百分率较低，差异有统计学意义（P〈0．05）。结论采用微量进样器钻孔较磨钻钻孔能有效减轻移植损伤，明显提高NSCs存活率，有利于神经元生长，并减少胶质瘢痕形成。Objective To compare the influence of two different neural stem cells （NSCs） transplantation methods in brain injury and graft sustain in mice. Methods NSCs were separated from C57BL/6 mice embryos and labeled using 5 - bromine deoxidization uracil nucleoside （Brdu）. Two methods were utilized to implant NSCs suspension （5×10^5 cells, 5 μL） into the different cerebral hemispheres of the same strain adult mice （n=42）： the skull was penetrated by a drill on the left side and a syringe needle on the right. Animals were randomly sacrificed on days 1, 2, 3, 7, 14, 21 and 28 of transplantation, and the brain sections were stained for morphology analysis, survival of grafts （HE） and distribution of neurons and astrocytes （Brdu/neuronal nuclear antigen [NeuN] and Brdu/glial fibrillary acidic protein [GFAP] double labeling immunofluorescence）. Results （1） As compared with that by the syringe needle skull penetration （the right hemisphere）, the brain structure was obviously damaged by the drill approach （the left hemisphere）; on days 1 and 3 of implantation, wide-spread necrosis of resident cells, spillover of red blood cells and infiltration of inflammatory cells were detected in the left hemisphere; 7 days after implantation, visible scar tissue formed and gathered more and more in the transplantation area until 28 days in the left hemisphere. （2） The distribution of Brdu labeled NSCs was relatively concentrated on days 1 and 3 of implantation, and the migration was obviously observed on days 7-21; besides, the percentage of Brdu positive cells on the left side was significantly lower than that on the right in each observation point （P〈0.05）. （3） Transplanted NSCs gave raise to neurons and astrocytes; more astrocytes and fewer neurons were detected in the left hemisphere, and the differences of NeuN and GFAP positive cells between two hemispheres had statistical significance （P〈0.05）. Conclusion In contrast to the drill penetration, the syringe needle skull penet

关 键 词：神经干细胞 移植 脑损伤 神经元 星形胶质细胞 

分 类 号：R651.1[医药卫生—外科学]