Cardiomyocyte Regeneration from Multipotent precursor c*

描述（由申请人提供）： 这项研究的目的是开发技术，促进心脏，肺和其他类型的细胞从体干细胞再生。 尽管已知多能细胞存在于许多成人组织中，但其有限的增殖和分化能力限制了其用于再生疗法。 通过对调节分化的因子的分子遗传学水平的了解，将有助于改善对靶组织的贡献的功效。 迄今为止，体干细胞的分化主要在组织损伤的整个动物模型中进行评估。 由于整个动物模型不容易有利于发现调节分化的基因和蛋白质，因此本申请提出了四个目的来设计和使用体外培养测定以鉴定体干细胞分化的分子调节剂。 重点将放在，但不限于，心肌细胞分化。 前两个目标将设计培养试验，以评估有希望的体干细胞群（称为SP细胞）的分化，该细胞群可以从一系列分化组织（包括心脏）中富集FACS。 目的1将评估促进胚胎心肌发生的因素和组织是否刺激这些细胞分化。初步数据描述了种间组织重组试验，以评估体外分化。 类似地，目标2将检验分化心肌提供促进分化的因子的假设。 目的3和4是设计用于鉴定合成的和天然的分化调节剂的中到高通量测定。 目标3将定义体干细胞向心肌细胞分化过程的分子标记物，这些标记物将用作这些测定的终点。 目的4建立小分子筛选机理并进行中试筛选. 为了与其他目标同时实现目标4，最初将使用胚胎干细胞，因为它们在培养中增殖良好，并响应于已知的诱导剂分化为心肌细胞（初步数据）。

DESCRIPTION (provided by applicant): The objective of the research is to develop technology that will facilitate the regeneration of heart, lung, and other cell types from somatic stem cells. Although multipotent cells are known to reside within numerous adult tissues, their limited proliferative and differentiative capacities constrain their use for regenerative therapies. Prospects for improving efficacy of contribution to target tissue will be facilitated by molecular genetic-level knowledge of the factors that regulate differentiation. To date, differentiation of somatic stem cells has been evaluated primarily in whole animal models of tissue damage. Since whole animal models are not readily conducive for the discovery of genes and proteins that regulate differentiation, this application proposes four aims to design and use in vitro, culture assays to identify molecular regulators of somatic stem cell differentiation. Emphasis will be placed on, but not limited, to cardiomyocyte differentiation. The first two aims will design culture assays to evaluate differentiation of a promising somatic stem cell population (known as SP cells) that can be FACS enriched from a range of differentiated tissues, including heart. Aim 1 will evaluate whether factors and tissues that promote embryonic cardiomyogenesis stimulate these cells to differentiate. Preliminary data describe interspecies tissue recombination assays to evaluate differentiation in vitro. Similarly, aim 2 will test the hypothesis that differentiated myocardium provides factors that promote differentation. Aims 3 and 4 are to design moderate to high throughput assays for the identification of synthetic and natural modulators of differentiation. Aim 3 will define molecular markers of the somatic stem cell to cardiomyocyte differentiation process that will be used as endpoints in these assays. Aim 4 will establish the mechanics of small molecule screens and carry out pilot screens. To carry out Aim 4 concurrently with the other aims, embryonic stem cells will be used initially as they proliferate well in culture and differentiate into cardiomyocytes in response to known inducers (preliminary data).