软骨来源成体干细胞的分离及其鉴定

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (4): 412-417.

软骨来源成体干细胞的分离及其鉴定

左伟¹,吴志宏²,苏新林¹,吴南¹,邱贵兴¹

1. 中国医学科学院北京协和医学院北京协和医院
2. 北京协和医院

收稿日期:2012-10-25 修回日期:2012-12-31 出版日期:2013-04-05 发布日期:2013-03-15
通讯作者: 邱贵兴 E-mail:qiuguixingpumch@gmail.com
基金资助:
国家自然科学基金

Isolation and identification of adult stem cells from articular cartilage

Received:2012-10-25 Revised:2012-12-31 Online:2013-04-05 Published:2013-03-15

摘要/Abstract

摘要： 目的观察关节软骨中是否存在着软骨干细胞，并对软骨干细胞进行分离、培养和鉴定。方法将软骨组织消化成单个软骨细胞，接种到预先用纤连蛋白处理过的平皿中，20mins后给予换液，培养2周时用克隆环挑取形成的单个细胞集落，在体外进行扩增。流式细胞仪检测扩增后细胞表面标志物，并对细胞的成脂肪、成骨、成软骨能力进行验证。结果成功挑选出软骨细胞集落，培养至第3代，细胞呈均一的长梭形。细胞表面表达间充质干细胞相关抗原CD44、CD29、CD73、CD90和CD166，且几乎不表达CD45、CD34和CD133。当给与诱导培养基培养时，软骨干细胞能成功的向脂肪细胞、骨细胞和软骨细胞进行分化。结论关节软骨中存在着软骨干细胞，并成功的对软骨干细胞进行了分离和鉴定。

关键词: 软骨, 干细胞, 修复

Abstract: Objective To observe whether there are mesenchymal stem cells in the articular cartilage, and identify the cartilage-derived stem cells. Methods The isolated single chondrocytes digested from cartilage tissue were seeded onto the fibronectin coated plates .After 20 mins, media and non-adherent cells were removed, fresh media were added to the remaining adherent cells. 2 weeks after plating , colonies were selected and isolated using sterile cloning rings. Then we detected the surface markers with flow cytometry and verified the ability of adipogenic, osteogenic and chondrogenic differentiations of the cells. Results The morphological feature of the third passage’s cartilage-derived stem cells displayed an uniform fibroblast-like phenotype. The cells were positive for many stem cells markers, including CD44、CD29、CD73、CD90 、CD166, and negative for CD45 、CD34and CD133,as shown by flow cytometry. Adipogenic, osteogenic and chondrogenic differentiations were revealed in cartilage-derived stem cells when cultured in specific medium． Conclusion There are mesenchymal stem cells in human articular cartilage, and we have isolated and identified the cartilage-derived stem cells successfully.

Key words: articular cartilage , stem cells , repair

左伟吴志宏苏新林吴南邱贵兴. 软骨来源成体干细胞的分离及其鉴定[J]. 基础医学与临床, 2013, 33(4): 412-417.

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