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目的观察体外培养外周血内皮祖细胞(EPCs)增殖、迁移和黏附能力与EPCs端粒酶活性之间的关系。方法密度梯度离心法获取人外周血单个核细胞(MNCs),接种至明胶包被的培养板上,培养7d后进行细胞免疫组化鉴定EPCs,采用MTT比色法、Transwell小室和黏附能力测定实验,观察EPCs的增殖、迁移和黏附能力。TRAP-PCR银染法检测EPCs端粒酶活性。结果从人外周血能成功分离EPCs细胞。从冠心病患者分离的EPCs增殖、迁移、黏附能力较非冠心病患者下降。冠心病患者EPCs端粒酶活性较非冠心病患者低。结论冠心病患者EPCs功能显著减弱,可能与EPCs端粒酶活性降低有关。
Objective To observe the relationship between the proliferation, migration and adhesion of EPCs cultured in vitro and the telomerase activity of EPCs. Methods Human peripheral blood mononuclear cells (MNCs) were obtained by density gradient centrifugation. The cells were seeded onto gelatin-coated plates and cultured for 7 days. EPCs were identified by MTT colorimetric assay, Transwell chamber assay and adhesion assay , Observe EPCs proliferation, migration and adhesion. The telomerase activity of EPCs was detected by TRAP-PCR silver staining. Results EPCs were successfully isolated from human peripheral blood. EPCs isolated from patients with coronary heart disease proliferation, migration, adhesion decreased compared with non-coronary heart disease patients. The telomerase activity of EPCs in patients with coronary heart disease was lower than that in non-coronary patients. Conclusion The function of EPCs in patients with coronary heart disease was significantly weakened, which may be related to the decrease of telomerase activity in EPCs.