Brunt Keith R, Hall Sean R R, Ward Christopher A, Melo Luis G
Department of Physiology, Queen's University, Ontario, Canada.
Methods Mol Med. 2007;139:197-210. doi: 10.1007/978-1-59745-571-8_12.
Endothelial progenitor cells (EPCs) and mesenchymal stem cells (MSCs) have emerged as potentially useful substrates for neovascularization and tissue repair and bioengineering. EPCs are a heterogeneous group of endothelial cell precursors originating in the hematopoietic compartment of the bone marrow. MSCs are a rare population of fibroblast-like cells derived from the bone marrow stroma, constituting approximately 0.001-0.01% of the nucleated cells in the marrow. Both cells types have been isolated from the bone marrow. In addition, EPC can be isolated from peripheral blood as well as the spleen, and MSC has also been isolated from peripheral adipose tissue. Several approaches have been used for the isolation of EPC and MSC, including density centrifugation and magnetic bead selection. Phenotypic characterization of both cell types is carried out using immunohistochemical detection and fluorescence-activated cell sorting analysis of cell-surface molecule expression. However, the lack of specific markers for each cell type renders their characterization difficult and ambiguous. In this chapter, we describe the methods that we use routinely for isolation, characterization, and genetic modification of EPC and MSC from human, rabbit, and mouse peripheral blood and bone marrow.
内皮祖细胞（EPCs）和间充质干细胞（MSCs）已成为用于血管新生、组织修复和生物工程的潜在有用细胞底物。EPCs是起源于骨髓造血区室的一组异质性内皮细胞前体。MSCs是源自骨髓基质的一类罕见的成纤维细胞样细胞，约占骨髓中有核细胞的0.001 - 0.01%。这两种细胞类型均已从骨髓中分离出来。此外，EPC可从外周血以及脾脏中分离得到，而MSC也已从外周脂肪组织中分离出来。已采用多种方法分离EPC和MSC，包括密度离心和磁珠分选。使用免疫组织化学检测和细胞表面分子表达的荧光激活细胞分选分析对这两种细胞类型进行表型鉴定。然而，每种细胞类型缺乏特异性标志物使得它们的鉴定困难且不明确。在本章中，我们描述了我们常规用于从人、兔和小鼠外周血及骨髓中分离、鉴定和基因改造EPC和MSC的方法。
