成人骨髓间充质干细胞体外向视网膜细胞的诱导分化  

Human bone marrow-derived mesenchymal stem cells differentiate into retinal cells in vitro

作　　者：俞海燕[1] 吴文涛[1] 王薇[2] 张纯[1] 

机构地区：[1]北京大学第三医院临床干细胞研究中心,北京100191 [2]北京大学第三医院眼科中心,北京100191

出　　处：《中国药理学通报》2014年第6期787-791,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30170999);北京市科委资助项目(No H020220010390)

摘　　要：目的研究取材于成人骨髓的间充质干细胞在一定诱导条件下向视网膜神经细胞的分化。方法成人骨髓经密度梯度离心得到的细胞,根据其高黏附特性体外培养获得间充质干细胞。利用流式细胞仪分析其细胞表型,在体外诱导使其向视网膜神经细胞分化并用免疫荧光法进行鉴定。结果从骨髓中分离培养的细胞具有成纤维细胞样形态,贴壁生长,表型相对均一,表面标志为CD90、CD44、CD147阳性;而CD34、CD38、CD45、CD14、HLA-DR阴性。体外诱导后可以得到表达nestin(神经干细胞标志物)、GFAP(神经胶质细胞标志物)和Rhodopsin(视网膜光感受器细胞标志物)阳性的细胞。结论从人骨髓中分离培养得到的间充质干细胞具有向视网膜神经细胞分化的潜能。Aim To study the differentiation of human bone marrow-derived mesenchymal stem cells （ HM-SCs） into retinal cells in vitro. Methods HMSCs were isolated from human bone marrow after Ficoll den-sity gradient centrifugation. The adherent cells after at least 5 passages were used for study. Immunopheno-type of the cells was analysed by flow cytometer, and cellular differentiation was identified by immunofluores-cence labeling technique. Results The target cells derived from human bone marrow adhered to the plate with fibroblastic-like morphology, whose surface mark-ers were similar to mesenchymal stem cells. Major cells 〈br〉 were positive for CD90 , CD44 , CD147 , while they were all negative for CD34, CD45, HLA-DR. In the differentiation study, HMSCs cultured in induced medium can differentiate into nestin （ neural stem cell ） -positive cell, GFAP （ glial fibrillary acidic protein ） -positive glial cells and retina-specific neurons express-ing Rhodopsin with CD90 （ mesenchymal stem cells ）-negative. Conclusion HMSCs have the ability to differentiate into retinal neural cells in vitro.

关 键 词：骨髓 间充质干细胞 视网膜 体外诱导分化 免疫荧光染色 RHODOPSIN 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学] R322.91[医药卫生—基础医学]