网孔过滤法分离纯化肌源性干细胞

Purification and Segregation of Rat Muscle-derived Stem Cells by Millipore Nylor Net Filters

探讨乳大鼠肌源性干细胞的体外培养及纯化分离方法。采用Ⅺ型胶原酶和胰蛋白酶消化乳大鼠骨骼肌,网孔过滤纯化肌源性干细胞后,通过体外原代和传代培养,经生长曲线、细胞融合率等指标研究肌源性干细胞的增殖和分化能力,免疫细胞化学染色法进行鉴定。结果表明:通过上述方法分离得到小圆形或短梭形的细胞,免疫细胞化学染色desmin及CD34为阳性。说明网孔过滤法可提高肌源性干细胞的纯度。

To establish the methods for culture, purification and segregation of rat muscle-derived stem cells in vitro. By two-step method, the rat muscle was digested with type XI collagen and trypsin, and the isolated cell suspension was purified by the millipore nylon net filter. The proliferation ability of MDSCs were analyzed through studying growth curve, and the obtained cells were identified with cell immunocytochemical technique. The results showed that the isolated MDSCs were small-cycloid or short-fusiform-shaped. They showed desmin^（＋） and CD34^（＋） by cell immunocytochemical technique. It proves that the method can improve the purification of the MDSCs through the millipore nylon net filters.