Characterization of Islet1+ Progenitors Derived from Human Embryonic Stem Cells

来源于出生后和胚胎心脏的多潜能Isl1+心血管前体细胞(MICPs)具有分化为心脏发生所需的关键细胞系：心肌细胞、血管内皮细胞和心肌细胞的能力。它们在成人心脏中的识别对心脏的恢复功能具有巨大的意义，但在分子水平上对这些前体细胞及其作用机制知之甚少。 指定的。我们最近开发了一种方法，通过这种方法，人胚胎干细胞(HESCs)可以在化学定义的培养液中统一分化为Isl1+祖细胞。这是一个很好的机会，可以彻底了解Isl1+祖细胞的基本生物学，也许可以将它们发展为潜在的细胞来源，最终可能用于心血管细胞治疗和组织工程。 这项建议将： 1.鉴定指定hESC来源的Isl1+前体细胞所需的信号通路。由于本规范需要WntSa和BMP4，因此将特别强调它们在Isl1+前体细胞生成中的作用。 2.为Isl1+祖细胞的维持和扩增创造条件。这将包括实验，其中研究支撑Isl1+状态的分子基础，包括分析信号通路 下游转录因子网络对Isl1+祖细胞的发育很重要。 3.假设人胚胎干细胞来源的Isl1+前体细胞具有向心肌细胞、平滑肌细胞和内皮细胞分化的潜能，建立其分化能力。同时，还有一种能力 将评估Isl1+祖细胞在心脏和后肢缺血动物模型系统中的功能。 我们开发的统一、强大的Isl1+祖细胞生成方法有可能显著提高我们对与心脏发育相关的早期胚胎事件的理解，以及与心血管疾病相关的细胞疗法的发展。

Multipotent Isl1+ cardiovascular precursors (MICPs) derived from post-natal and embryonic hearts have the capacity to differentiate into the critical cell lineages required for cardiogenesis: cardiomyocytes, smooth muscle and endothelial cells. Their identification in the adult heart has enormous implications for cardiac restorative function but little is known about these progenitors at the molecular level and how they are specified. We have recently developed a method whereby human embryonic stem cells (hESCs) can be uniformly differentiated into Isl1+ progenitors in chemically defined media. This represents an excellent opportunity to thoroughly understand the basic biology of Isl1+ progenitors and perhaps, to develop them as a potential source of cells that could eventually be used for cardiovascular cell therapies and tissue engineering. This proposal will: 1. characterize signaling pathways required for specification of hESC-derived Isl1+ progenitors. Since WntSa and BMP4 are required for this specification, special emphasis will be placed on their role in the generation of Isl1+ progenitors. 2. establish conditions for maintenance and amplification of Isl1+ progenitors. This will involve experiments where the molecular basis underpinning the Isl1+ state is investigated including analysis of signaling pathways and downstream transcription factor networks important for Isl1+ progenitor development. 3. establish the differentiation capacity of hESC-derived Isl1+ progenitors with the hypothesis that they have the potential to generate cardiomyocytes, smooth muscle cells and endothelial cells. In parallel, the ability of Isl1+ progenitors to function in animal model systems for cardiac and hind limb ischemia will be evaluated. The uniform, robust method for generation of Isl1+ progenitors that we have developed has the potential to significantly advance our understanding of early embryonic events associated with cardiac development and, for the development of cell therapeutics associated with cardiovascular disease.