人外周血内皮祖细胞免疫磁珠分离、纯化及鉴定的实验研究
首发时间:2012-11-13
摘要:目的 从人外周血中用免疫磁珠分离内皮祖细胞,考察其细胞形态和纯度,并对其进行内皮祖细胞功能的鉴定。方法 以CD133免疫磁珠从人外周血中分离出内皮祖细胞,通过光学显微镜观察细胞形态,并通过流式细胞仪对细胞的表型进行了分析,用共聚焦显微镜对内皮祖细胞的功能进行了鉴定。结果 从人外周血中免疫磁珠分离得到的内皮祖细胞CD133+细胞比例在97%以上,细胞形态良好,并具有内皮祖细胞结合FITC标记的荆豆凝聚素1和吞噬DiI标记的低密度脂蛋白的功能。结论 从人外周血中免疫磁珠分离可得到高纯度的内皮祖细胞并表现出内皮祖细胞的功能,是组织工程、基因与细胞治疗理想的载体。
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Isolation and identification of endothelial progenitor cells from human peripheral blood by immunomagnetic beads
Abstract:Objective To isolate and identify endothelial progenitor cells (EPCs) from human peripheral blood by immunomagnetic beads, and to study the cell morphology, purification and function. Methods: CD133+ immunomagnetic beads were used to isolate EPCs from peripheral blood. The morphology of EPCs were observed by optical microscopy. The purification of EPCs was investigated by flow cytometry. The fuction of EPCs was characterzied by confocal laser scanning microscope. Results: CD133+ EPCs were successfully isolated from human peripheral blood by immunomagnetic beads. The percentage of CD133 positive cells is more than 97% of the isolated cells. The endothelial-lineage characteristics of expanded cells were confirmed by FITC-UEA-1 binding and DiI-ac-LDL uptake assay with the aid of confocal laser scanning microscope. Conclusion Highly purified EPCs from human peripheral blood by immunomagnetic beads showed the endothelial-lineage characteristics, which would be an ideal carrier for tissue engineering, gene and cell therapy.
Keywords: endothelial progenitor cells huaman peripheral blood immunomagnetic beads
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