胎儿骨髓间质干细胞的获得和诱导成软骨培养  

Isolation of fetal mesenchymal stem cells and induced chondrocyte culture

作　　者：庞永刚[1] 崔鹏程[1] 陈文弦[1] 

机构地区：[1]解放军第四军医大学唐都医院耳鼻咽喉科,陕西省西安市710038

出　　处：《中国临床康复》2003年第29期3934-3935,T002,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金(30171007)~~

摘　　要：目的:研究胎儿骨髓间质干细胞(MSCs)体外分离、培养、扩增并向软骨细胞表型转化,探究新的组织工程化软骨种子细胞的来源。方法:从引产胎儿(家属自愿捐献)髂骨获得骨髓,体外培养从骨髓中获得的MSCs,以流式细胞仪检测细胞表面抗原。将传代后的MSCs在诱导培养液中培养,检测软骨细胞的特异性标志物。结果:从胎儿骨髓得到的细胞表达MSCs的表面标志物,并可在体外培养、扩增,诱导后细胞变为多伪足的多边形细胞,扫描电镜见细胞表面有丰富的交联,透射电镜观察见大量粗面内质网、高尔基体、线粒体。甲苯胺蓝染色见细胞内有异染性基质(免疫组化和RT-PCR检测COLⅡ表达阳性。结论:胎儿MSCs可作为软骨组织工程的较理想的种子细胞。AIM:To study the isolation, culture, expansion and chondrocytic differentiation of the fetal mesenchymal stem cells(MSCs) in vitro and investigate new source of tissue engineer cartilage cell. METHODS:MSCs were isolated from the fetal bone marrow(family contribute voluntarily).The cells surface antigens of the fetal MSCs were detected by flow cytometry and MSCs were cultured in the special medium to induce the chondrocytic differentiation.Specific markers were detected in cartilage cell.RESULTS:Flow cytometry showed that the cells derived from fetal bone marrow expressed the markers of MSCs and the cells can be cultured and expanded in vitro.The cell became multi pseudopodium polygon cell.There were plentiful adherence on cell surface by scanning electron microscope. There also were mass rough endoplasmic reticulum, Golgi apparatus and mitochondrion.It had metachromasia stroma by using toluidine blue staining and COLⅡ expressed positive by immunochemistry and RT PCR.Fetal MSCs cultured in the medium expressed the markers of chondrocytes.

关 键 词：胎儿 骨髓间质干细胞 软骨细胞 MSCS 免疫组化 

分 类 号：R336[医药卫生—人体生理学]