Defining Therapeutic Potential of Clonal Stem Cell Populations using Targeted Nan

项目摘要 胚胎干细胞为细胞替代疗法提供了相当大的潜力 因受伤或疾病而丧失，因为他们有能力持续自我更新和区分 几乎进入任何类型的细胞或组织。然而，目前人类胚胎干细胞的过程 细胞(HESCs)分化为各种成熟的功能细胞类型还知之甚少，存在 对扩增hESCs并将其分化为所需细胞类型的方法的严重需求，如 如上皮细胞、真皮细胞、内皮细胞、心肌细胞和骨骼肌细胞等。 技术已经分离出数百个克隆细胞群(称为谱系限制细胞，LRC) 源于分化hESCs，这些hESCs似乎既可在培养中扩张，又具有 多种成熟细胞类型的早期祖细胞的特性。我们想研究LRC 在人类胚胎干细胞接收信号的差异化文化背景下的差异 以更好地了解它们的治疗潜力以及如何引导其 文化中的差异化。近期，我们的目标是识别针对LRC的特定靶向多肽 噬菌体展示。对这些同质祖代群体的选择将使我们能够识别特定的 靶向以表面标记为靶标的多肽，否则在以下情况下可能会被低估 在分化的hESCs的异源群体上进行选择。我们将追踪发展的命运 利用量子点共轭区分hESCs的天然环境中靶向LRCs的研究 靶向多肽。这些工具将允许使用研究特定的祖细胞的分化 使用延时成像实时发生。用2个或更多的多肽进行多重标记将使我们能够 人胚胎干细胞体外分化过程中祖细胞间的相互作用。这些研究将被用来 筛选具有较高治疗潜力的喉癌细胞，探索喉癌的分化方法。我们的龙 学期目标是开发精选的LRC，用于皮肤、血管和心脏的再生治疗 以及LRCs及其作为研究试剂的多肽靶向剂的商业化。 。项目叙事 胚胎干细胞为细胞替代疗法提供了相当大的潜力 细胞因损伤或疾病而丧失了持续自我的能力 更新和分化为几乎任何类型的细胞或组织的能力。有一个关键的问题 人类胚胎干细胞体外扩增和分化为理想干细胞的方法尚未满足的需求 细胞类型，如皮肤，心肌，血管等。我们已经分离了数百个 来源于可在培养中扩张的人胚胎干细胞的纯血统限制细胞(LRC) 并具有多种成熟细胞类型的早期祖细胞的特性。附近 术语，我们的目标是用量子点标记靶向试剂来标记特定的LRC 我们可以在分化hESCs和hESCs的培养中追踪LRCs的发育命运 从而确定它们的治疗潜力。我们的长期目标是开发精选的 LRCs用于皮肤、血管和心脏等的再生治疗，如 以及LRCs及其多肽靶向剂的商业化研究 试剂。

Project Summary Embryonic stem cells offer considerable potential for cell replacement therapy where cells have been lost to injury or disease because of their capacity for continual self renewal and ability to differentiate into virtually any cell type or tissue. Currently, however the process by which human embryonic stem cells (hESCs) differentiate into various mature functional cell types is poorly understood and there is a critical unmet need for methods of expanding and differentiating hESCs into desirable cell types such as epithelial, dermal, endothelial, cardiac and skeletal myocytes, etc. Scientists at Advanced Cell Technology have isolated hundreds of clonal cell populations (called lineage restricted cells, LRCs) derived from differentiating hESCs which appear to be both expandable in culture and have the properties of early progenitor cells of a multitude of mature cell types. We would like to study LRC differentiation in the context of a differentiating culture of hESCs where they receive signals from surrounding cells and matrix to better understand their therapeutic potential and how to direct their differentiation in culture. Near term, our goal is to identify specific targeting peptides for LRCs using phage display. Selection on these homogeneous progenitor populations will allow us to identify specific targeting peptides that target surface markers that might otherwise be under-represented when selecting on a hererogeneous population of differentiating hESCs. We will track the developmental fate of targeted LRCs in their native context of differentiating hESCs using quantum dot conjugated targeting peptides. These tools will allow use to study the differentiation of specific progenitor cells as it occurs in real time using time-lapse imaging. Multiplex labeling with 2 or more peptides will allow us to trace progenitor cell interactions during in vitro differentiation on hESCs. These studies will be used to select LRCs with high therapeutic potential and to develop methods for LRC differentiation. Our long term goal is to develop selected LRCs for use in regenerative therapies for skin, vasculature, and heart as well as commercialization of the LRCs and their peptide targeting agents as research reagents. . Project Narrative Embryonic stem cells offer considerable potential for cell replacement therapy where cells have been lost to injury or disease because of their capacity for continual self renewal and ability to differentiate into virtually any cell type or tissue. There is a critical unmet need for methods of expanding and differentiating hESCs in-vitro into desirable cell types such as skin, heart muscle, blood vessels, etc. We have isolated hundreds of pure lineage restricted cells (LRCs) derived from hESCs which are expandable in culture and have the properties of early progenitor cells of a multitude of mature cell types. Near term, our goal is to label specific LRCs with quantum dot labeled targeting agents so that we can track the developmental fate of LRCs in cultures of differentiating hESCs and thus determine their therapeutic potential. Our long term goal is to develop selected LRCs for use in regenerative therapies for skin, vasculature, and heart among others as well as commercialization of the LRCs and their peptide targeting agents as research reagents.