人脂肪间充质干细胞的分离、培养及鉴定

Isolation,Cultivation and Identification of Human Adipose-derived Stem Cells

取脂肪吸脂术分离的脂肪细胞,在胰酶和胶原酶的作用下分离、培养ADSC并对其进行增殖能力、干细胞表面标志的测定以及转录因子水平的检测.分离、培养的ADSC在光镜下呈圆形、贴壁生长,可见小的胞浆突起.经细胞增殖曲线绘制,细胞在1～3d处于生长适应期,而3～6d处于对数生长期,8～10d后细胞增殖明显减缓.流式细胞术鉴定细胞表面标志结果显示分离培养的细胞表达间充质干细胞表面标志,而不表达造血干细胞和成纤维细胞表面标志.对培养的ADSC检测其转录因子Nanog和oct-4的表达,结果显示体外培养的ADSC在第15代仍可稳定表达Nanog和oct-4.

ADSCs were isolated from human subcutaneous adipose tissue acquired from people who underwent elective liposuction. We detected the proliferation capacity of the ADSCs, and then analyzed ADSCs surface markers profile by flow cytometry. The expression levels of transcription factors were detected by RT PCR. The results of flow cytometry identification show that ADSCs expressed the surface markers of bone marrow stem cells instead of expressing the surface markers of hematopoietic stem cells or of fibrocytes. In culturing period of passage 15, ADSCs also stably expressed Nanog and oct-4.