目的研究分离、培养小鼠骨髓血管内皮祖细胞的方法并对其进行诱导分化鉴定,为临床进行缺血性疾病的替代治疗奠定细胞学基础。方法用灌注法分离小鼠骨髓血管内皮祖细胞,用血管内皮生长因子、成纤维细胞生长因子诱导其分化,形态学观察细胞的生长过程,并利用抗CD34抗体鉴定血管内皮祖细胞,利用抗vWF抗体、抗eNOS抗体鉴定血管内皮细胞。结果小鼠骨髓血管内皮祖细胞可在体外培养条件下贴壁生长,可诱导分化为表达特异性组织蛋白的内皮细胞。结论在小鼠骨髓内可分离出骨髓源性血管内皮祖细胞,呈贴壁增殖状态,并有分化能力。
Objective To study the method of isolation,cultivation of endothelial progenitor cells from BALB/c mice bone marrow and identification of its differentiation,to establish cytological foundation for vicariousness therapy of ischemic disease.Methods The endothelial progenitor cells were isolated by injector perfusion method from bone marrow of BALB/c mice,and induced by VEGF and bFGF,morphology method was used to observe cells growth status,anti-CD34 antibody was used to test the specificity of endothelial progenitor cells,anti-vWF antibody and anti-eNOS antibody were used to identify cell's differentiation by immunocytochemistry test.Results The status of endothelial progenitor cell from BALB/c mice bone marrow was adherence culture in vitro,and it was induced by cytokine to differentiate endothelial cell with typical morphologic and expressing tissue-specific protein.Conclusions The endothelial progenitor cell can be isolated from BALB/c mice with adherent proliferation status and has potential to differentiation.