Wisconsin Stem Cell Research Center

描述(申请人提供)：我们最近从恒河猴、普通绒猴和人类获得的胚胎干细胞系对人类发育生物学、药物发现、药物测试和移植医学具有广泛的意义。在培养皿中培养数月或数年后，这些细胞保持了形成从心肌到神经到血液--可能是构成身体的任何细胞类型--的细胞的能力。人类ES细胞的增殖和发育潜力有望为体外实验研究和心脏病、帕金森病和白血病等疾病的移植治疗提供基本无限的特定细胞类型。然而，从长远来看，人类胚胎干细胞最大的遗产可能不是作为移植医学的组织来源，而是作为了解人体的基础研究工具。这一新工具对于人类和小鼠之间明显不同的血统来说将特别重要。这里提出的人类胚胎干细胞研究探索性中心将开始专注于利用人类胚胎干细胞来研究基础人类生物学中其他方法不容易解决的问题。一个中心核心将提供人类ES细胞组织培养支持，包括培养液准备、质量控制、成纤维细胞准备以及对培养的人类ES细胞进行常规的Sky染色体分析。在资助的最后一年，将组织一个小型的、有重点的研讨会，将人类胚胎干细胞研究人员和其他基础生物学家聚集在一起，以确定人类胚胎干细胞领域的挑战和机会。我们提出了以下试点计划： (I)我们将分析一个重要的早期谱系：从人类ES细胞到原始外胚层到神经外胚层到分化神经元的转变。(Ii)我们将分析BMP4如何导致人类ES细胞退出自我更新并分化为滋养层细胞的转录调控，使用表达克隆策略和iRNA来增加和减少功能。(Iii)我们将使用生化方法，结合尖端的质谱学和蛋白质组学，识别那些由成纤维细胞产生的调节人类ES细胞自我更新的因子。

DESCRIPTION (provided by applicant): Our recent derivation of embryonic stem (ES) cell lines from rhesus monkeys, common marmosets, and humans has widespread implications for human developmental biology, drug discovery, drug testing, and transplantation medicine. After months or years of growth in culture dishes, these cells retain the ability to form cells ranging from heart muscle to nerve to blood--potentially any cell type that makes up the body. The proliferative and developmental potential of human ES cells promises an essentially unlimited supply of specific cell types for in vitro experimental studies and for transplantation therapies for diseases such as heart disease, Parkinson's disease, and leukemia. In the long run, however, the greatest legacy for human ES cells may be not as a source of tissue for transplantation medicine, but as a basic research tool to understand the human body. This new tool will be particularly important for lineages that differ significantly between humans and mice. The Exploratory Center for Human Embryonic Stem Cell Research proposed here will begin to focus on using human ES cells to study questions in basic human biology that are not easily addressed by other approaches. A center core will provide human ES cell tissue culture support, including media preparation, quality control, fibroblast preparation, and routine SKY chromosomal analysis of cultured human ES cells. In the final year of funding, a small, focused workshop will be organized to bring together human ES cell researchers and other basic biologists to identify challenges and opportunities in the human ES cell field. We propose the following Pilot Projects: (I) We will analyze the transciptional changes in one important early lineage: the transition from human ES cell to primitive ectoderm to neural ectoderm to differentiated neurons. (II) We will analyze the transcriptional control of how BMP4 causes human ES cells to exit self-renewal and differentiate to trophoblast, using expression cloning strategies and iRNA to add and subtract function. (III) We will use a biochemical approach, coupled with cutting-edge mass spectrometry and proteomics, to identify those factors produced by fibroblasts that mediate human ES cell self-renewal.