成人骨髓间质干细胞向神经干细胞的诱导分化  

Induced differentiation of adult human BMSCs into neural stem cells

作　　者：庄琴[1] 朱彦[1] 陆益龙[1] 唐爱国[1] 宗海洋[1] 贺清华[2] 陈谦[2] 姜平[2] 张志坚[2] 

机构地区：[1]江苏大学附属医院血液科,江苏镇江212001 [2]江苏大学基础医学与医学技术学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2011年第2期93-96,F0002,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(30570981)

摘　　要：目的:建立诱导成人骨髓间质干细胞(bone marrow mesenchymal stem cells,BMSCs)向神经干细胞分化的体外培养方法,为细胞移植修复神经系统损伤提供种子细胞。方法:原代贴壁培养成人BMSCs并传代培养至第3代时,将细胞接种于铺有琼脂糖的培养瓶,用神经干细胞培养基(含B27,EGF,bFGF)抗贴壁诱导培养。将诱导培养的神经球(neurospheres)传代扩增。用免疫荧光染色和蛋白质印迹法,对贴壁培养的BMSCs和抗贴壁诱导培养的神经球细胞表达神经干细胞标志蛋白的水平进行检测和比较。结果:抗贴壁诱导培养可使BMSCs形成神经球,初级神经球传代可扩增形成次级神经球。神经球细胞高表达音猬因子、神经细胞黏附分子以及神经干细胞标志蛋白巢蛋白和CD133。结论:琼脂糖抗贴壁,神经干细胞培养基诱导培养可使成人BMSCs形成神经球,神经球细胞具有神经干细胞样特征,该方法可为细胞移植修复神经系统损伤提供大量的种子细胞。Objective： To establish an in vitro method to induce the differentiation of adult human bone marrow mesenchymal stem cells（BMSCs） into neural stem cells as the seed cells for the transplant therapy of neural system injuries.Methods： Human BMSCs were isolated from bone marrow and cultured for three passages（P3）.The BMSCs were plated to the agarose-coated anti-adherent flasks,using the neural stem cell culturing medium including B27,EGF and bFGF to induce the differentiation of BMSCs to neurospheres.With the passage of neurospheres,neural stem cells（NSCs） proliferated.At last the NSCs markers in adherent cultured BMSCs and anti-adherent plus inducing cultured cells grew as neurospheres were compared with immunofluorescence and Western blotting.Results： Anti-adherent plus inducing culture can derive neurospheres from BMSCs.The primary neurospheres can be passaged to secondary neurospheres with proliferation.The cells in neurospheres expressed sonic hedgehog,neural cell adhesion molecule（NCAM）,as well as the neural stem cell markers Nestin and CD133.Conclusion： The method combining anti-adherent with agarose and NSCs culturing medium can induce the differentiation of adult human BMSCs into neurospheres with NSCs characteristics,and is one promising strategy to provide large quantities of seed cells for repairing neural system injuries.

关 键 词：骨髓间质干细胞 神经干细胞 巢蛋白 CD133 神经细胞黏附分子 音猬因子 琼脂糖 

分 类 号：R329.1[医药卫生—人体解剖和组织胚胎学]