人骨髓间充质干细胞在新生小鼠脑中的植入和分化

Engraftment and Differentiation of Human Bone Marrow Mesenchymal Stem Cells in Neonatal Mouse Brain

背景与目的:探讨人骨髓间充质干细胞(humanmesenchymalstemcells,hMSCs)在体内向神经细胞分化的可能性。材料与方法:将标记绿色荧光蛋白的人骨髓间充质干细胞植入到新生3d的小鼠侧脑室中,分别于移植后0d,9d和14d处死受体鼠,取其脑组织,4%多聚甲醛固定后行冠状面冰冻切片,荧光显微镜下观察hMSCs的植入,激光共聚焦显微镜检测植入细胞神经特异性蛋白的表达,定位双重标记的植入细胞。结果:受体小鼠脑中均可检出植入的细胞,此类细胞表达神经元细胞特异的微管相关蛋白β-Ⅲ-Tubulin(Tuj1)、微管相关蛋白2(MAP2),一些细胞表达神经胶质细胞特异的胶质原纤维酸性蛋白(GFAP)。结论:hMSCs植入后受到脑组织特定微环境的影响,在体内可以向神经细胞分化并参与到发育的神经系统中。

BACKGROUND ＆ AIM：To explore the possibility of human mesenchymal stem cells （hMSCs） differentiating into neural cells in vivo. MATERIALS AND METHODS： hMSCs labeled with enhanced green fluorescent protein （EGFP） were transplanted into the lateral ventricle of neonatal mouse brain. At 0, 9 and 14 days post-transplantation, mice were sacrificed and their brains were fixed with 4% paraformaldehyde. The engraftment of transplanted cells in coronal section of the recipient mouse brain was examined under a fluorescent microscope. Indirected immunofluorescent staining was used to detect the expression of neural proteins of these grafted cells, RESULTS： Examination of cryostat section under fluorescent microscopy revealed that hMSCs could engraft into neonatal mouse brain. Neuronal differentiation was confirmed by the expression of β-Ⅲ-Tubulin （Tujl） and MAP2 of these EGFP-expressing cells, whilst some grafted cells had GFAP staining. CONCLUSION： These results suggest that hMSCs could respond to brain microenvironment and differentiate into neural cells.