Isolation of Perivascular Multipotent Precursor Cell Populations from Human Cardiac Tissue.
Isolation of Perivascular Multipotent Precursor Cell Populations from Human Cardiac Tissue.
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DOI：
10.3791/54252
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发表时间：
2016-10
期刊：
影响因子：
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通讯作者：
J. Baily;William C W Chen;N. Khan;I. Murray;Z. N. Gonzalez Galofre;J. Huard;B. Péault
中科院分区：
文献类型：
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作者：
J. Baily;William C W Chen;N. Khan;I. Murray;Z. N. Gonzalez Galofre;J. Huard;B. Péault
多能间充质干细胞/基质细胞（MSC）通常通过其塑料粘附从原代组织消化物中分离出来，但其解剖组织位置仍不清楚。我们小组和其他研究人员最近发现了多种组织中血管周围细胞明确的血管周围和间充质干细胞标记物表达，这为前瞻性分离和纯化多能血管周围前体细胞的特定同质亚群提供了机会。我们之前已经演示了使用荧光激活细胞分选 (FACS) 从人类骨骼肌中纯化微血管 CD146+CD34- 周细胞和血管 CD34+CD146- 外膜细胞。在此，我们描述了一种通过 FACS 从人类心肌中同时分离这两种血管周围细胞亚群的方法，该方法基于一组定义的细胞表面标记物的表达，用于阳性和阴性选择。因此，该方法使得多能心脏 MSC 样前体细胞的两个特定亚群可用于基础研究和/或治疗研究。
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 CD146+CD34- pericytes and vascular CD34+CD146- 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.