摘要
目的 :探讨体内外环境对骨髓间充质干细胞 (MSCs)向神经细胞分化的影响。方法 :①DAPI体外标记MSCs后 ,直接注入大鼠大脑中动脉梗塞 (MACO)模型 (n =6 )的脑内 ,2周后取脑组织 ,用免疫荧光检测MSCs来源细胞的巢蛋白 (nestin)、微管相关蛋白 - 2 (MAP - 2 )、胶质纤维酸性蛋白 (GFAP)的表达。②体外用全反式视黄酸 (ATRA)预诱导 2 4h后 ,换用改良的神经细胞培养基 (MNM )培养 18h ,免疫组化检测nestin、MAP - 2、GFAP的表达。结果 :①植入 2周后 ,MSCs在注射部位及其周围广泛分布 ,仅有少许细胞表达nestin、MAP - 2和GFAP。②MSCs在体外诱导后 ,nestin、MAP - 2的阳性率分别为 (92 .3± 3.4 ) %和 (89.6 +3.3) % ,但不表达GFAP。结论 :MSCs在脑内向神经细胞转化的转化率较低 ,而在体外转化为神经元的转化率相当高 ,体内外环境对MSCs向神经细胞的分化的过程及机理存在较大差异。
Objective:To investigate the effects of environment in vitro and in vivo on the differentiation of bone mesenchymal stem cells (MSCs) into neural cells.Methods:①MSCs were labeled with 4,6-diamidino-2-phenylindole (DAPI) and injected into the right brain of the rats(n=6),which had suffered from middle cerebral artery occlusion(MACO) a day before.Two weeks later,immunofluorescent staining was used to detect the expression of nestin,microtubule-associated protein 2 (MAP-2) and glial fibrillary acidic protein (GFAP).②In vitro,MSCs were cultured with modified neuronal medium(MNM)for 18 hours after being pretreated with 0.5umol/L all-trans retinoid acid (ATRA) for 24 hours.Immunocytochemistry was used to detect the expression of nestin,MAP-2 and GFAP.Results:①After being injected into rats for 2 weeks,MSCs survived in the ischemic areas and a few cells expressed nestin,MAP-2 and GFAP respectively.②In vitro,after ATRA and MNM treatment,the positive percentage of nestin and MAP-2 were (92.3±3.4)% and (89.6±3.3)% respectively,but no expression of GFAP was detected.Conclusion:The different ratio of MSCs differentiating into neural cells suggests that there is difference in the process and mechanism between environment in vitro and in vivo.
出处
《重庆医科大学学报》
CAS
CSCD
2004年第5期588-590,628,共4页
Journal of Chongqing Medical University
基金
国家自然科学基金(NO :3 0 0 70 3 82 )
重庆市科委科学基金(渝科委计(2 0 0 1) 5 4号文 )
重庆市卫生局科学基金(渝卫科教(2 0 0 1) 46号文 )
关键词
骨髓间充质干细胞
神经细胞
分化
Mesenchymal Stem Cells
Neural cells
Differentiation