肿瘤患者循环血管内皮干/祖细胞的检测及体外诱导分化

目的　证实外周血中循环血管内皮细胞 (CECs)及血管内皮前体细胞 (CEPs)的存在 ,检测内皮前体细胞的增殖分化能力。方法　应用流式细胞术检测 5 7例肿瘤患者及 15例正常人外周血中的CECs和CEPs ,应用ELISA方法检测血清VEGF水平。选择经造血干细胞动员的肿瘤患者 ,分离单个核细胞 (MNCs) ,在VEGF、IGF及bFGF条件下培养血管内皮细胞 ,并应用透射电镜法对培养细胞进行鉴定。结果　肿瘤患者外周血中CECs、CEPs含量分别为 0 .3 78%± 0 .0 47%、0 .0 5 9%± 0 .0 13 % ,高于正常对照组 ,且与血清VEGF水平相关。透射电镜下培养细胞可见Weibel-Palade小体 ,为血管内皮细胞的特异性标志。结论　外周血中存在CECs和CEPs ,在肿瘤患者体内存在血管内皮及其前体细胞的动员 ,循环血管内皮干 /祖细胞可进一步分化成熟

Detection and induced differentiation of Circulating Endothelial Cells and Progenitors in Peripheral Blood in Patient with Tumor

Objective To testify the presence of circulating endothelial cells(CECs) and circulating endothelial precursors(CEPs) in peripheral blood and their ability of proliferation and differentiation.Methods CECs and CEPs were measured in 57 patients with tumor and 15 healthy controls by 3-color flow cytometry.The serum level of VEGF was determined by ELISA method.Freshly isolated mononuclear cells(MNCs) from G-CSF mobilized peripheral blood were cultured in the presence of VEGF,IGF and bFGF.Cultured cells were identified by immunohistochemistry(IHC) and transmission electron microscopy(TEM).Results CECs/CEPs in patients with tumor were significantly increased.The amount of CECs/CEPs was positively correlated with serum level of VEGF.TEM studies in cultured cells showed structures compatible with Weibel-Palade bodies that are found exclusively in vascular endothelium.Conclusion We testify the existing of CECs and CEPs in peripheral blood.The amount of CECs/CEPs increased in patients with tumor.The endothelial precursors can differentiate into mature endothelial cells.