Marking Progenitor Cell Lineages in the Vascular System

描述(由申请人提供)： 干细胞是一种自我更新的多能细胞，可以产生多种细胞谱系和/或身体的细胞类型。人和小鼠胚胎干细胞的发展以及成体干细胞的分离受到了人们的广泛关注。使用干细胞/祖细胞来替代受损或患病的人体组织的可能性已经得到了广泛的宣传。这项技术的一个应用将是修复受损的心脏和血管组织；然而，这将首先需要鉴定和表征血管干细胞/祖细胞。该项目的目标是鉴定和鉴定血管干/祖细胞，并探索其在体外和体内的发育潜力。我们将用一种适合于流式细胞术的自动荧光报告程序来标记早期的血管前体细胞，例如增强型绿色荧光蛋白(EGFP)。我们将在小鼠胚胎干细胞中的早期内皮基因的内源性基因中“敲入”自动荧光标签。其次，我们将从分化的ES细胞和胚胎卵黄囊膜中分离和鉴定有标记的祖细胞群体。第三，我们将在体外和体内检测这些祖细胞的谱系分化潜力。我们期望这个项目将提供重要的洞察，在血管系统的开始干细胞的性质，并可能共同的造血细胞和内皮细胞谱系。最终，这些研究将解决使用干细胞治疗受损或患病血管的基本问题。

DESCRIPTION (provided by applicant): Stem cells are self-renewing, pluripotent cells that can give rise to multiple cell lineages and/or cell types of the body. The development of human and mouse embryonic stem cells and the isolation of adult stem cells have received much attention. The possibility of using stem/progenitor cells to replace damaged or diseased human tissue has been widely publicized. One application of this technology would be to repair damaged cardiac and vascular tissue; however, this will first require identification and characterization of vascular stem/progenitor cells. The goal of this project is to identify and characterize vascular stem/progenitor cells and to explore their developmental potential in vitro and in vivo. We will mark early vascular progenitor cells with an autofluorescent reporter suitable for flow cytometry, such as enhanced green fluorescent protein (EGFP). We will "knock-in" the auto fluorescent tag into the endogenous locus of early endothelial genes in mouse embryonic stem (ES) cells. Second, we will isolate and characterize marked progenitor populations from differentiating ES cells and embryonic yolk sac membranes. Third, we will examine the lineage differentiation potential of these progenitor cells in vitro and in vivo. We anticipate that this project will provide important insights into the nature of stem cells at the beginning of the vascular system, and possibly common to the hematopoietic and endothelial lineages. Ultimately, these studies will address basic questions of using stem cells for damaged or diseased blood vessels.