摘要
目的研究冠心病患者外周血中内皮祖细胞的数量、形态和活性。方法选择冠心病患者57例和正常对照者30例,用密度梯度离心法从外周血获取单个核细胞,将其接种在人纤维连接蛋白包被的培养板,用加入血管内皮生长因子165和碱性成纤维细胞生长因子的1640培养基培养细胞,并分析细胞形态和形成集落的数量,7d后贴壁细胞进行细胞分析和计数。用激光共聚焦显微镜鉴定FITC标记的荆豆凝集素和Dil标记的乙酰化低密度脂蛋白,双染色阳性细胞为正在分化的内皮祖细胞;用流式细胞仪检测细胞表面抗原CD34和KDR;采用MTT比色法检测细胞的生长状态。结果冠心病患者外周血内皮祖细胞数量较对照组明显减少(23.1±1.8个/200倍比56.7±2.4个/200倍,P<0.05),形成细胞集落数(14.7±2.5个/40倍比24.2±1.7个/40倍,P<0.05)、细胞增殖能力和生长曲线也明显降低。结论冠心病患者外周血内皮祖细胞的数量和活性明显降低。
Aim To investigate the isolation and culture of endothelial progenitor cells （EPC） from peripheral blood in patients with coronary heart diseases （CHD）, and to determine cell shape, amount, activity and clusters, Methods Mononuclear cells were isolated from peripheral blood of patients with CHD by Ficoll-density contrifugation. The isolated cells were cultured in 1640 medium supplemented with vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （ 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 marked acetylated low density lipoprotein （acLDL-Dil） uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPC proliferation were assayed by MTT as-say. Results The number of EPC was significantly reduced in patients with CHD compared with control subjects （23.1±1.8 vs 56.7±2.4, P 〈 0.05 ）, and the number of cell clusters was also reduced in patients with CHD compared with control subjects （ 14.7±2.5 vs 24.2±1.7, P〈0.05）. The proliferation was also impaired in patients with CHD. Conclusion EPC were cultured and differentiated at specific conditions derived from mononuclear ceils of peripheral blood.
出处
《中国动脉硬化杂志》
CAS
CSCD
2008年第5期381-384,共4页
Chinese Journal of Arteriosclerosis
基金
湖南省教育厅"十一五"重点项目(2006-56)
关键词
内科学
内皮祖细胞
冠心病
细胞数量
集落
增殖
Endothelial Progenitor Cells
Coronary Heart Disease
Cell Number
Ousters
Proliferation
