脑缺血条件下骨髓基质细胞分化成少突胶质细胞的研究

The differentiation of mesenchymal stem cells into oligodendrocyte under cerebral ischemia

目的观察体外培养的人胚胎骨髓基质细胞(MSCs)移植入全脑缺血大鼠胼胝体内能否定向分化成少突胶质谱系细胞。方法无菌人胚胎股骨骨髓体外分离培养,应用免疫荧光流式细胞仪鉴定MSCs细胞表型,MSCs用B rdu标记后在立体定向仪下移植入脑缺血大鼠胼胝体内,用免疫荧光双染色及激光共聚焦显微镜观察B rdU-A2B5、B rdU-O4、B rdU-CNPase双阳性表达情况。结果传代3次的人胚MSCs CD44、CD29阳性表达细胞占95%以上,而CD34阳性表达细胞仅0.14%;移植点附近可见B rdU标记的MSCs移植入脑缺血大鼠胼胝体内存活。免疫荧光双染色显示,部分B rdU阳性细胞表达O4,但无B rdU-A2B5、B rdU-CNPase双阳性细胞。结论人胚MSCs移植入脑缺血大鼠胼胝体内能部分定向分化成少突胶质谱系细胞,但分化延迟。

Objective To investigate whether the cultured mesenchymal stem cells （MSCs） derived from bone marrow of human aborted fetal could differentiate into oligodendrocyte after transplanted into ischemic corpus callosum. Methods MSCs of human aborted fetal were isolated and cultured aseptically. Immunofluore scence staining was used to identify MSCs phenotype by flow cytometer analysis. MSCs labeled with BrdU were transplanted into ischemic corpus callosum with the aid ofa stereotaxie instrument. Immunofluorescence double staining was used to identify BrdU-A2BS,BrdU-O4,BrdU-CNPase double positive cells under confocal microscopy. Results Over 95% of the passage 3 human MSCs were CD44/CD29 positive cells with only 0.14% of CD34 positive cells. The BrdU-labeled MSCs which were transplanted into ischemic corpus callosum survived around the transplant point. A portion of BrdU positive cells expressed O4 simultaneously. None of them expressed A2B5 and CNPase simultaneously by immunofluorescence double staining. Conclusion Few human MSCs which were transplanted into ischemic corpnscallosum could differentiate into oligodendrocyte lineage cells, but it deferred.