Embryonic Stem Cell Derived Cardiac Myocytes

该研究领域涉及胚胎干细胞分化为心肌细胞之前和期间的研究。在这些研究中，我们使用胚胎干（ES）细胞（R1，D3）、胚胎生殖细胞（EG-1）和胚胎癌细胞（P19）。我们已经建立了一个最有效的系统，可用于从ES细胞在体外产生心肌细胞，我们已经建立了当ryanodine受体，SR钙ATP酶，受磷蛋白和二氢吡啶受体首次表达与体外分化，允许EC耦合与分化的研究。选择协议（最近与心脏限制部分的Na/Ca交换启动子）已允许心肌细胞的分离。总的来说，该研究旨在产生心脏谱系特异性细胞，以了解调节蛋白在体外心肌细胞形成中的作用。此外，如果细胞可以被分离到同质性，并在体外分化后显示出活力，那么我们建议使用这种技术来测试啮齿动物中基于细胞的治疗后的递送方案和心脏功能的改善。这一基础研究工作的一个主要重点是致力于提高这些细胞在体外培养条件下的活力。干细胞中的几个靶基因也被修饰，以确定它们在心肌细胞分化和存活中的作用，最近，我们已经开始分离（和靶向）细胞，以选择可能更适合基于细胞的治疗的心脏祖细胞亚群。通过研究胚胎干细胞的基本生物学，我们希望描绘出负责心肌细胞发育和更新的机制，并将这些结果应用于改善可能适用于人类的基于细胞的疗法。

SUMMARY OF WORK This research area involves the study of embryonic stem cells prior to and during differentiation to cardiomyocytes. For these studies, we employ embryonic stem (ES) cells (R1, D3), embryonic germ cells (EG-1) and embryonic carcinoma cells (P19). We have established one of the most efficient systems available for the generation of cardiomyocytes from ES cells in vitro, and we have established when the ryanodine receptor, SR CaATPase, phospholamban and dihydropyridine receptor were first expressed with in vitro differentiation, permitting studies of EC coupling with differentiation. Selection protocols (most recently with a cardiac-restricted portion of the Na/Ca exchanger promoter) have permitted the isolation of cardiomyocytes. Overall, the research is aimed at generating cardiac-lineage specific cells to understand the role of regulatory proteins in the formation of cardiomyocytes in vitro. Additionally, if the cells can be isolated to homogeneity and shown to be viable after in vitro differentation, then we propose using this technique to test delivery protocols and improvements of cardiac function following cellular based therapies in rodent. A major focus of this basic research effort is devoted to the improved viability of these cells during in vitro cultivation conditions. Several target genes are also being modified in the stem cells to determine their role in cardiomyocyte differentiation and survival, and more recently, we have begun isolating (and targeting) cells to select sub-populations of cardiac progenitor cells that may be more appropriate for cellular based therapies. By studying the basic biology of embryonic stem cells, we hope to delineate mechanisms responsible for cardiomyocyte development and renewal, and apply these results to improve cellular based therapies that may be applicable to man.