【摘要】 目的　分离纯化人骨髓间质干细胞 (MSC) ,研究其基本生物学特性。方法　用比密 1 .0 73Ficoll淋巴细胞分离液分离成人骨髓MSC ,体外扩增 ,观察细胞生长特性 ,流式细胞仪检测骨髓MSC表面抗原表达及细胞周期 ,染色体技术分析骨髓MSC遗传特性。结果　成人骨髓MSC培养 3d后有散在呈针尖状的贴壁细胞 ,7～ 1 0d后形成集落或融合呈纤维状 ;体外扩增原代可获得 (4～ 6 )× 1 0 5个细胞 ,1 0代可获得 (1～ 5 )× 1 0 1 0 个细胞 ,5～ 6代以后的细胞增殖能力下降 ;流式细胞仪检测结果显示 :CD1 3、CD2 9、CD4 4、CD71表达阳性 ,CD3、CDl4、CD33、CD34、CD38、CD4 5、HLA DR不表达 ;染色体核型正常 ;细胞周期分析显示 ,G0 /G1 期 :79.4 8% ,G2 /M期 :6 .1 0 % ,S期 :1 4 .4 2 %。结论　人骨髓MSC体外具有较好的增殖更新能力 ,3～ 6代的人骨髓MSC作为组织工程细胞具有广阔的应用前景更多还原

【Abstract】 Objective To isolate human mesenchymal stem cell (MSC) from adult bone marrow and investigate its basic biological characteristics.Methods Human MSC was separated from adult human marrow by specific gravity 1.073 Ficoll reagent and expanded in culture medium.The proliferation and growth characteristics were observed in primary and passage culture.The surface antigens and cell cycle of human MSC were analysed by flow cytometer. The karyotype of human MSC was detected.Results The isolated human MSC comprised a single phenotypic population and displayed a fibroblast like morphology.Human MSC had a strong self renewal capacity.Three days later,MSC began to adhere to culture bottle and displayed spindle shape in culture.After ten days,it began to fuse as fibroblast like.After primary culture,approximately(4～6)×10 5 cells were obtained.After passage 10,(1～5)×10 10 cells were got.The proliferative ability decreased gradually after five or six passages.These expanded attached human MSC was positive for CD13?CD29?CD44?CD71 but did not express CD14?CD33?CD34?CD38?CD45?HLA DR.Karyotype of human MSC was normal.Analysis for cell cycle included G 0/G 1 phase:79.48%,G 2/M phase:6.10%,S phase:14.42%.Conclusion Human MSC has strong self renewal capacity and can be used as the seed cells for tissue engineering during passages3～6 in vitro.更多还原