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Isolation of Endothelial Progenitor Cells from Human Umbilical Cord Blood

机译:从人脐带血中分离内皮祖细胞

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摘要

The existence of endothelial progenitor cells (EPCs) in peripheral blood and its involvement in vasculogenesis was first reported by Ashara and colleagues. Later, others documented the existence of similar types of EPCs originating from bone marrow,. More recently, Yoder and Ingram showed that EPCs derived from umbilical cord blood had a higher proliferative potential compared to ones isolated from adult peripheral blood,,. Apart from being involved in postnatal vasculogenesis, EPCs have also shown promise as a cell source for creating tissue-engineered vascular and heart valve constructs,. Various isolation protocols exist, some of which involve the cell sorting of mononuclear cells (MNCs) derived from the sources mentioned earlier with the help of endothelial and hematopoietic markers, or culturing these MNCs with specialized endothelial growth medium, or a combination of these techniques. Here, we present a protocol for the isolation and culture of EPCs using specialized endothelial medium supplemented with growth factors, without the use of immunosorting, followed by the characterization of the isolated cells using Western blotting and immunostaining.
机译:Ashara及其同事首次报道了外周血中内皮祖细胞(EPC)的存在及其与血管生成的关系。后来,其他人记录了源自骨髓的类似类型的EPC的存在。最近,Yoder和Ingram显示,与从成人外周血中分离出的EPC相比,从脐带血中提取的EPC具有更高的增殖潜力。除了参与产后血管生成外,EPCs还显示出有望作为创建组织工程化血管和心脏瓣膜构建体的细胞来源。存在多种分离方案,其中一些方案涉及借助内皮和造血标记物对源自前述来源的单核细胞(MNC)进行细胞分选,或将这些MNC与专门的内皮生长培养基一起培养,或将这些技术结合使用。在这里,我们提出了一种协议,用于在不使用免疫分选的情况下使用添加了生长因子的专用内皮细胞培养基进行EPC的分离和培养,然后使用Western印迹和免疫染色对分离的细胞进行表征。

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