摘要
目的为梗死心肌促血管新生的实验研究提供方法和移植细胞的来源。方法利用密度梯度离心法分离人外周血单个核细胞,在内皮环境中培养并鉴定。CD31+/vWF+细胞被认为是所需要的内皮祖细胞。MTT实验观察贴壁细胞的增殖活性。结果培养的细胞符合EPC的形态学特征、细胞表面标志和功能特征。培养3d和5d,CD133+细胞分别占贴壁细胞的2.6%和9.3%。培养7d每毫升外周血可获得(2.7±1.2)×105个贴壁细胞,其中CD31+/vWF+细胞占54.2%。培养第5天到第7天以及第9天传代后2d与第3天的贴壁细胞相比,细胞增殖活性有显著的统计学意义(P<0.001)。结论利用外周血能够培养出以内皮祖细胞为主的细胞群。
Objective To provide methods and cell resource for therapeutic neovascularization of myocardial infarction. Methods Mononuclear cells from human peripheral blood were isolated using a density-gradient centrifugation method and incubated in M199 medium containing 20% fetal calf serum and 4 mg/L bovine pituitary extract. Markers for endothelial cells were identified using flow cytometry system. MTT assay was used to study the activity of proliferation. The morphological characteristics of attaching cells were also studied. Results Positive expressions of CD133 at day 4 and day 6 were 2.6% and 9. 3% of total adherent cells,respectively. After 7 d of culture,the cells showed the appearance of cell clusters, spindle-shaped and attaching cells, and cord-like structures. 54.2% ± 8.6% of total adherent cells were CD31^＋/vWF^＋ double positive by flow cytometry analysis. There were significant differences in the proliferation activity from day 5 to day 9 compared with that of day 3 in culture with MTT assay （ P 〈0. 001 ）. Conclusion Endothelial progenitor cells can be isolated from peripheral blood and expanded in vitro.
出处
《安徽医科大学学报》
CAS
北大核心
2005年第6期519-522,共4页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金安科医药专项资助项目(编号:01043711)
关键词
内皮细胞
内皮
血管/细胞学
干细胞
细胞培养
endothelial cells
endothlium,vascular/cytology
stem cells
cell culture
