用VEGF165和bFGF诱导单个核细胞分化成内皮祖细胞的试验研究

目的研究从健康人外周血中提取单个核细胞，用VEGF165和bFGF诱导细胞分化成内皮祖细胞，并分析细胞形态、成集落数量，细胞数量活性。方法选择健康人20例，用密度梯度离心法从外周血获取单个核细胞，用生长因子VEGF165和bFGF诱导细胞分化，7d后将贴壁细胞进行分析。荧光显微镜鉴定FITC-UEA-I和DiI-acLDL双染色阳性细胞为正在分化的EPC；流式细胞仪检测CD34和KDR；MTT法检测细胞生长状态；并分析生长因子VEGF165和bFGF干预后细胞形态和成集落的数量。结果VEGF165和bFGF干预后EPC的细胞数目和集落数明显增加，细胞增殖活性明显增强。结论成人外周血中提取的单个核细胞，在VEGF165和bFGF作用下可培养为内皮祖细胞。

Mononuclear cells were induced into endothelia progenitor cells by VEGF165 and bFGF

Objective To determine the biological traits and optimal condition for the induction and differentiation of endothelial progenitor cells from peripheral blood in healthy adults. Methods Mononuelear cells were isolated from peripheral blood of healthy adults by Ficoll-density eentrifugation. The isolated ceils were cultured in 1640 medium supplemented with VECF165 and bFGF. The EPC specific surface mark CD34 and KDR were assessed by fluorescence activated cell sorter(FACS)analysis: EPC were characterized as adherent cells double positive for DiL-acLDL uptake and lectin binding by direct fluorescent staining under a hser scanning confocal microscope. EPC migration were assayed by MTr assay. Result The number and migration ability of EPC were increased by VEGFl65 and bFGF. Conclusion Endothelial progenitors cells can be derived from mononuclear cells of peripheral blood at specific conditions.