Baily James E, Chen William C W, Khan Nusrat, Murray Iain R, González Galofre Zaniah N, Huard Johnny, Péault Bruno
Centre for Cardiovascular Science, Queen's Medical Research Institute, University of Edinburgh.
Department of Bioengineering and Orthopaedic Surgery, University of Pittsburgh; Research Laboratory of Electronics and Department of Biological Engineering, Massachusetts Institute of Technology;
J Vis Exp. 2016 Oct 8(116):54252. doi: 10.3791/54252.
Multipotent mesenchymal stem/stromal cells (MSC) were conventionally isolated, through their plastic adherence, from primary tissue digests whilst their anatomical tissue location remained unclear. The recent discovery of defined perivascular and MSC cell marker expression by perivascular cells in multiple tissues by our group and other researchers has provided an opportunity to prospectively isolate and purify specific homogenous subpopulations of multipotent perivascular precursor cells. We have previously demonstrated the use of fluorescent activated cell sorting (FACS) to purify microvascular CD146CD34 pericytes and vascular CD34CD146 adventitial cells from human skeletal muscle. Herein we describe a method to simultaneously isolate these two perivascular cell subsets from human myocardium by FACS, based on the expression of a defined set of cell surface markers for positive and negative selections. This method thus makes available two specific subpopulations of multipotent cardiac MSC-like precursor cells for use in basic research and/or therapeutic investigations.
多能间充质干细胞（MSC）传统上是通过其贴壁特性从原代组织消化物中分离出来的，但其解剖学组织位置尚不清楚。我们团队和其他研究人员最近发现，多种组织中的周血管细胞表达特定的周血管和MSC细胞标志物，这为前瞻性分离和纯化多能周血管前体细胞的特定同质亚群提供了机会。我们之前已经证明了使用荧光激活细胞分选（FACS）从人骨骼肌中纯化微血管CD146CD34周细胞和血管CD34CD146外膜细胞。在此，我们描述了一种基于用于阳性和阴性选择的一组特定细胞表面标志物的表达，通过FACS从人心肌中同时分离这两个周血管细胞亚群的方法。因此，该方法可提供两种特定的多能心脏MSC样前体细胞亚群，用于基础研究和/或治疗研究。
