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联合应用胶原酶消化、贴壁筛选及单克隆培养法从人脂肪组织中分离、培养人脂肪干细胞(hADSCs),观察hADSCs的形态学特征。通过流式细胞术检测hADSCs表面抗原;通过细胞化学染色和免疫荧光观察hADSCs的一般形态学特征;利用透射电镜分析hADSCs的超微结构。结果显示体外分离、培养出高度同源性的hADSCs,hADSCs具有相对特异的细胞表面标志,即CD29、CD44、CD90、CD105及CD166呈阳性表达,而CD31、CD45及HLA-DR呈阴性表达;hADSCs苏木精-伊红染色见胞浆淡染,油红O及碱性磷酸酶(AKP)染色阴性;免疫荧光CD29、CD166呈阳性;透射电镜下可见hADSCs细胞质内粗面内质网、线粒体丰富,细胞表面可见丰富微绒毛。本实验结果为hADSCs在细胞、组织再生移植修复中的应用提供了形态学基础。
Human adipose tissue stem cells (hADSCs) were isolated and cultured from human adipose tissue with collagenase digestion, adherent screening and monoclonal culture. Morphological characteristics of hADSCs were observed. The surface antigens of hADSCs were detected by flow cytometry. The morphological features of hADSCs were observed by cytochemical staining and immunofluorescence. The ultrastructure of hADSCs was analyzed by transmission electron microscopy. The results showed that highly homologous hADSCs were isolated in vitro. HADSCs had relatively specific cell surface markers, ie, positive expression of CD29, CD44, CD90, CD105 and CD166, but negative expression of CD31, CD45 and HLA-DR. Hematoxylin-eosin staining showed that the cells were stained lightly, and oily red O and alkaline phosphatase (AKP) were negative. The immunofluorescence staining of CD29 and CD166 was positive. The transmission electron microscopy showed that the rough endoplasmic reticulum in hADSCs was cytoplasmic, , The surface of the cell can be seen rich microvilli. The results of this study provide a morphological basis for the application of hADSCs in the repair of cell and tissue regeneration grafts.