Embryonic Stem Cell Derived Cardiac Myocytes

胚胎干细胞衍生的心肌细胞

• 批准号: 7592067
• 负责人: Kenneth R Boheler
• 金额: $ 93.53万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7592067
• 关键词: Area; Basic Science; Biology; Cardiac; Cardiac Myocytes; Cell Survival; Cells; Condition; Coupling; Development; Dihydropyridine Receptors; ES Cell Line; Embryo; Gene Targeting; Generations; Germ Cells; Goals; In Vitro; Laboratories; Malignant Epithelial Cell; Mus; Population; Protocols documentation; Research; Research Personnel; Rodent; Role; Ryanodine Receptor Calcium Release Channel; Stem cells; System; Techniques; Testing; Therapeutic; Work; base; embryonic stem cell; genetic regulatory protein; improved; man; phospholamban; promoter; Area; Basic Science; Biology; Cardiac; Cardiac Myocytes; Cell Survival; Cells; Condition; Coupling; Development; Dihydropyridine Receptors; ES Cell Line; Embryo; Gene Targeting; Generations; Germ Cells; Goals; In Vitro; Laboratories; Malignant Epithelial Cell; Mus; Population; Protocols documentation; Research; Research Personnel; Rodent; Role; Ryanodine Receptor Calcium Release Channel; Stem cells; System; Techniques; Testing; Therapeutic; Work; base; embryonic stem cell; genetic regulatory protein; improved; man; phospholamban; promoter

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.

工作总结该研究领域涉及在分化为心肌细胞之前和分化过程中胚胎干细胞的研究。对于这些研究，我们采用胚胎干细胞（R1，D3），胚胎生殖细胞（EG-1）和胚胎癌细胞（P19）。我们已经建立了一种最有效的系统之一，用于从ES体外从ES细胞产生心肌细胞，并且当Ryanodine受体，SR CAATPase，Phospholamban和Dihydropylidine受体首先用体外分化表达，从而与分化相结合的EC耦合研究时，我们已经建立了。选择方案（最近具有NA/CA交换器启动子的心脏限制部分）允许分离心肌细胞。总体而言，该研究旨在产生心脏差的特定细胞，以了解调节蛋白在体外形成心肌细胞中的作用。此外，如果可以将细胞分离为均匀性并在体外区分后证明是可行的，那么我们建议使用该技术在啮齿动物中基于细胞的疗法后使用该技术测试递送方案和改善心脏功能。这项基础研究工作的主要重点是在体外培养条件下这些细胞的生存能力提高。在干细胞中还修改了几种靶基因，以确定其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疗法的心脏祖细胞的亚群。通过研究胚胎干细胞的基本生物学，我们希望描述负责心肌生物发育和更新的机制，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。