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,20 mins后换液,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.用诱导培养基培养时,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.%Objective To confirm the presence of 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, os-teogenic 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,CD34 and CD133,as shown by flow cytometry. Adipogenic, osteogenic and chondrogenic differentiations were revealed in cartilage-derived stem cells when cultured in specific medium. Conclusions There are mesenchymal stem cells in human articular cartilage, and we have isolated and identified the cartilage-derived stem cells successfully.
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