外周血中循环血管内皮细胞及血管内皮前体细胞的扩增与鉴定

目的证实外周血中循环血管内皮细胞(circulating endothelial cells,CECs)及血管内皮前体细胞(circulating endothelial precursors,CEPs)的存在,以及内皮前体细胞的增殖分化能力.方法应用流式细胞法检测57例肿瘤患者及15例正常人外周血中的CECs/CEPs.选择经造血干细胞动员的肿瘤患者,分离单个核细胞(mononuclear cells,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0.378±0.047%、0.059±0.013%,高于正常对照组.培养细胞透射电镜下可见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/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.

Expansion and Identification of circulating endothelial cells and progenitors in peripheral Blood

Objective To testify the presence of circulating endothelial cells (CECs) and circulating endothelial precursors (CEPs) in the peripheral blood. Methods CECs and CEPs were enumerated in 57 tumor patients and 15 healthy controls by 3-color flow cytometry. 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 transmission electron microscopy (TEM). Results CECs/CEPs in tumor patients were significantly increased. TEM studies of cultured cells showed structures compatible with Weibel-Palade bodies that are found exclusively in vascular endothelium. Conclusion We testified the existence of CECs and CEPs in the peripheral blood. The amount of CECs/CEPs is increased in tumor patients. The endothelial precursors can differentiate into mature endothelial cells.