Ravishankar Prashanth, Zeballos M Alejandra, Balachandran Kartik
Department of Biomedical Engineering, University of Arkansas.
Department of Biomedical Engineering, University of Arkansas;
J Vis Exp. 2017 Sep 14(127):56021. doi: 10.3791/56021.
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.
阿沙拉及其同事首次报道了外周血中内皮祖细胞（EPCs）的存在及其在血管生成中的作用。后来，其他人记录了源自骨髓的类似类型EPCs的存在。最近，约德和英格拉姆表明，与从成人外周血中分离出的EPCs相比，源自脐带血的EPCs具有更高的增殖潜力。除了参与出生后的血管生成外，EPCs作为创建组织工程血管和心脏瓣膜构建体的细胞来源也显示出前景。存在各种分离方案，其中一些方案涉及借助内皮和造血标志物对源自上述来源的单核细胞（MNCs）进行细胞分选，或将这些MNCs与专门的内皮生长培养基一起培养，或结合这些技术。在这里，我们提出了一种使用补充有生长因子的专门内皮培养基分离和培养EPCs的方案，不使用免疫分选，然后使用蛋白质印迹和免疫染色对分离的细胞进行表征。
