人脂肪来源的间充质干细胞的生物学研究

目的探索从人脂肪中获取间充质干细胞的方法,通过观察人脂肪来源间充质干细胞(hADAS)的形态学、生长动力学、细胞表面标志,揭示其生物学特性。方法分离培养脂肪间充质干细胞并观察其形态学变化;四甲基偶氮唑盐(MTT)比色法测细胞活性;流式细胞仪测细胞周期和间充质干细胞表面抗原。结果脂肪干细胞呈成纤维细胞样;MTT比色法证实其有很强的增殖活性;流式细胞周期显示处于增生活跃期的细胞占很大比例;间充质干细胞表面抗原CD29、CD44随着传代表达逐渐增高,不表达内皮细胞标志CD31,造血干细胞标志CD34随细胞培养时间延长表达逐渐降低。结论通过酶消化法可从人脂肪中获取间充质干细胞,具有很强的增殖能力,表达干细胞表面标志。

Biological features of human adipose-derived adherent stromal cells

Objective To explore the methods for isolation and culcure of human adipose-derived adherent stromal ceils （hADAS） from adipose tissues, and to observe the morphology, cytodynamics and cell surface antigens of hADAS in order to reveal their biological features. Methods hADAS were isolated, cultured, and observed for morphology. The cell proliferation was assessed by MTT methods. Flow cytometry was performed for cell cycle analysis and expression of cell surface antigens. Results After 6 hours, the primarily cultured cells adhered to the wall. The cells spreaded and became fibroblast-like after culture for 12 hours. MTT assay showed highly active proliferation of hADAS cells. Flow cytometry showed that： stromal cell-associated markers （CD44, CD29） increased significantly with successive passages; endothelial cell-associated markers CD31 did not change significantly with serial passage;the stem cell- associated marker CD34 was at peak levels in the early-passage ASCs, although on the decline throughout the culture period, kept at a low level. Conclusion hADAS can be isolated from adipose tissue by enzymatic digestion and presented a fibroblast-like morphology. Proliferation of those cells were strongly active. Stromal cell surface antigens were found expressed on hADAS.