体外分离培养人脂肪间充质干细胞及鉴定

Isolation,cultivation and identification of human adipose- derived mesenchymal stem cells in vitro

目的建立体外分离培养人脂肪间充质干细胞(hADSCs)的方法。方法采用酶消化法分离培养hADSCs,流式细胞仪检测细胞免疫表型,台盼蓝染色计数绘制细胞生长曲线并计算倍增时间,油红和Von Kossa鉴定成脂成骨多向分化能力。结果成功从脂肪中分离纯化得到hADSCs,呈放射状、河流状的梭形细胞。流式结果显示hADSCs高表达CD29、CD44、CD73、CD90、CD105和CD166等,低表达或不表达CD45和HLA-DR。生长曲线和倍增时间显示自我更新及增殖能力强。成脂和成骨染色显示其有多向分化能力。结论实验表明建立了一种有效的hADSCs体外分离培养方法。

Objectine To establish the isolation,cultivation and identification methods of human adipose - derived mesenchymal stem cells（hADSCs）in vitro. Methods The hADSCs were isolated from abdomen epiploica adipose tissue by collagenase digestion. Expression of cel-lular markers,the proliferative capacity,and multipotential differentiation were respectively studied with flow cytometry analysis,cell counting,Oil Red O,and Von Kossa staining. Results The isolated hADSCs all displayed typical fibroblast - like morphological features. Phenotypic studies revealed that hADSCs were positive for CD29,CD44,CD73,CD90,CD105,CD166 and negative for CD45 and HLA - DR. The growth curve and doubling time analysis revealed high capability for self - renewal and proliferation. Oil Red O and Von Kossa staining demonstrated a multipo-tent differentiation capacity of hADSCs. Conclusion The data described here suggest that an efficient isolation and cultivation method of hADSCs has been established.