Cardiomyocyte Regeneration from Multipotent precursor c*

描述(由申请人提供)： 这项研究的目的是开发有助于从体细胞再生心、肺和其他类型细胞的技术。尽管已知多能细胞存在于许多成人组织中，但它们有限的增殖和分化能力限制了它们用于再生治疗。对调节分化的因素的分子遗传学水平的了解将有助于提高对靶组织的贡献的有效性的前景。到目前为止，体细胞的分化主要是在组织损伤的全动物模型中进行评估的。由于整个动物模型不利于发现调节分化的基因和蛋白质，本申请提出了四个目标，以设计和使用体外培养试验来识别体细胞分化的分子调节因子。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，以评估有希望的体细胞干细胞群体(称为SP细胞)的分化，这种干细胞可以从包括心脏在内的一系列分化组织中浓缩成FAC。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。初步数据描述了用于评估体外分化的种间组织重组试验。同样，目标2将检验分化的心肌提供促进分化的因素的假设。 目标3和4是设计中等到高通量的分析方法，用于鉴定合成的和天然的分化调节因子。AIM 3将定义体细胞到心肌细胞分化过程的分子标记物，这些标记物将被用作这些分析的终点。目标4将建立小分子筛分的机制，并进行中试筛选。为了同时实现目标4和其他目标，最初将使用胚胎干细胞，因为它们在培养中增殖良好，并在已知诱导剂(初步数据)的反应下分化为心肌细胞。

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).