小鼠骨髓来源内皮祖细胞体外诱导培养及分化特征

目的: 研究小鼠骨髓来源内皮祖细胞(EPCs)的体外培养方法及体外分化特征。方法: 采用梯度离心法分离小鼠骨髓单个核细胞并进行诱导培养,用流式细胞术和免疫细胞化学对细胞表型特征进行检测,分别采用MTT和Transwell方法对其增殖、迁移能力进行检测。结果: 体外培养4 d至14 d,CD133、SCA-1及CD117的表达率分别从11.05%、29.32%及16.45 %降至2.29%、7.82%及3.92%;而VEGFR-2、CD31、VE-cadherin、eNOS及vWF的表达率分别从12.21%、8.43%、18.27%、7.11%及32.61%增加至62.13%、32.96%、67.73%、27.89%及82.16%;吞噬acLDL和结合UEA-I的细胞从57.18%增加至86.70%。在0 μg/L至80 μg/L VEGF浓度梯度下,细胞增殖率和迁移率分别增加58.03%和33.83%。结论: EGM-2内皮培养基可稳定培养小鼠骨髓来源EPCs,培养过程中其干细胞标志物表达逐渐降低,而内皮细胞标志物表达及特性不断增加,VEGF可显著促进其增殖和迁移。

Characterization and differentiation of mouse bone marrow-derived endothelial progenitor cells

AIM:To investigate the culture method and differentiation characteristics of endothelial progenitor cells(EPCs) derived from mouse bone marrow.METHODS: Bone marrow mononuclear cells were isolated from BALB/c mice by density gradient centrifugation and cultured in fibronectin-coated dishes with EGM-2 medium.The phenotypic characteristics were determined by immunofluorescence and fluorescence-activated cell sorter.The proliferation and migration capacities were detected by MTT and Transwell assays,respectivel...