人脂肪源性干细胞体外培养和鉴定研究

Culture and Identification of Human Adipose Tissue Derived Stem Cells in Vitro

目的建立人脂肪源性干细胞分离、体外培养及鉴定方法,以期为组织研究和临床应用提供细胞来源。方法通过脂肪抽吸术获取人腹壁下脂肪组织,经过PBS液反复冲洗,I型胶原酶消化,采用直接贴壁筛选培养法进行体外培养,从脂肪组织中分离、培养、扩增细胞。相差显微镜观察细胞一般形态学特征,应用MTT法绘制细胞生长曲线,采用流式细胞仪检测细胞表面抗原,检测CD34、CD45、CD44、CD90表达。结果流式细胞仪检测表面标志物CD34、CD45表达阴性,CD44、CD90表达阳性。结论采用直接贴壁培养法进行体外培养可获得生长稳定、增殖能力强的脂肪干细胞。

Objective To establish a method for isolation,culture and identification of human adiposederived stem cell（ ADSCs）. Methods Abdominal adipose tissue was obtained from human through liposuction,washed with PBS,and then digested with type I collagenaseand cultured in vitro. The morphological characteristics of the cells were observed under a phase contrast microscope. MTT method was used to determine the growth curve and flow cytometry was adopted to detect the expression of surface antigens CD34,CD45,CD44 and CD90. Results The growth curve of human adipose-derived stem cells was opposite-like ＂S＂shape. The stem cells showed strong positive expression of surface markers CD44,CD90 and negative expression of CD34,CD45. Conclusion Direct adherent culture in vitro can obtain adipose tissue-derived steme cells with stable growth and strong proliferation ability.