人骨髓间充质干细胞的分离培养及其形态学特点  

Isolation,culture of human bone marrow mesenchymal stem cells and morphological characteristics

作　　者：何旭[1] 郭新[1] 崔丽[1] 王心蕊[1] 牛云[1] 李玉林[1] 

机构地区：[1]吉林大学病理生物学教育部重点实验室,吉林长春130021

出　　处：《电子显微学报》2006年第1期57-61,共5页Journal of Chinese Electron Microscopy Society

基　　金：863重大专项基金资助项目(No.2004AA205020);教育部博士点基金(No.20020183064);吉林大学青年教师基金(No.419070100050)

摘　　要：利用Percoll梯度分离、贴壁筛选法及单克隆培养法分离、培养人骨髓间充质干细胞(mesenchymal stem cells,MSCs);采用流式细胞术检测MSCs免疫学表型;采用细胞组织化学和免疫荧光技术观察MSCs的一般形态学特点;利用透射电镜和扫描电镜观察MSCs的超微结构。结果表明,分离、培养出高度同源性的MSCs具有独特的细胞免疫学表型,即CD73、CD105、CD44、CD166、CD90阳性,CD34、CD 31、CD45阴性;细胞组织化学MSCs PAS-过碘酸雪夫氏染色阳性,苏丹黑-B(SB)及碱性磷酸酶(AKP)染色阴性;免疫荧光见细胞CD73、CD105阳性;透射电镜下MSCs表面有微绒毛,相邻细胞间有缝隙连接,细胞器丰富;扫描电镜下MSCs呈长梭形、鱼群状排列,表面有许多微绒毛。实验结果为MSCs的诱导分化及应用提供了形态学基础。Human bone mesenchymal stem cells（MSCs） were isolated, cultured and proliferated by Percoll gradient centrifugation, adherence to plastic flask and monoclonal culture. Immunophenotypes of MSCs were detected by flow cytometry techniques. General morphological characteristics of MSCs were observed by cytochemical and immunofluorescent techniques. The uhrastructure of MSCs was observed by transmission and scanning electron microscopy. The results showed that high homogenous MSCs had unique immunophenotypes and they were positive for CD73, CD105, CD44, CD166 and CD90 but negative for CD34 ,CD31 and CD45. Cytochemistry showed that MSCs were positive for glycogen, and negative for SB and AKP. CD73 and CD105 were positive by immunofluorescence. There were abundant organella, microvilli and gap junction between two adjacent cells by transmission electron microscopy. MSCs showed long fusiform shape and flocked array by scanning electron microscopy. The results will offer a morphological foundation for differentiation and application of MSCs.

关 键 词：间充质干细胞 流式细胞术 细胞组织化学 免疫荧光 激光扫描共聚焦显微镜 超微结构 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]