Physiology of Neurons from Human & Mouse ES Cells

描述(申请人提供)：这项研究的目标是描述胚胎干细胞(ES)细胞来源的神经元的体外和体内生理学特征。这些细胞代表着用于研究和治疗的多潜能、遗传正常细胞的潜在无限来源。小鼠胚胎干细胞在体外可以分化为多种体细胞类型，包括神经元、星形胶质细胞和少突胶质细胞。此外，分化的ES细胞在移植到大脑或脊髓后存活，并在形态上与周围宿主组织整合。基于对小鼠ES细胞的研究，人类ES细胞的分离为新的替代疗法提供了可能性，在这种疗法中，体外分化的ES细胞将取代因损伤或疾病而失去的体细胞。神经系统的病理可能适合替代疗法，包括帕金森氏病、肌萎缩侧索硬化症、中风、亨廷顿病和多发性硬化症。布什总统决定允许联邦政府支持对现有人类胚胎干细胞系的研究，这激起了人们对探索干细胞来源的替代作为解决这些以前难以解决的缺陷的新方法的巨大热情；然而，在实现这种疗法之前，仍有许多基础研究要做。 我们的最终目标是开发将人类胚胎干细胞高效转化为特定类型神经元的程序，并在移植到宿主神经系统时优化胚胎干细胞来源的神经元与功能网络的整合。功能整合的一个重要组成部分是单个干细胞来源的神经元获得正常的生理特性。在这一点上，只有有限的信息可用关于分化的ES细胞的生理。因此，这项建议的目标是：1)评估来自不同ES诱导方案的神经细胞的生理分化。2)研究胚胎干细胞来源的神经元移植到脑内后的生理特性，包括与周围宿主神经元形成功能性突触连接的严格测试。人类和小鼠的胚胎干细胞将被并行使用，以比较它们的发展潜力。所有关于人类ES细胞的研究都将使用NIH人类ES细胞注册中的WA01品系。

DESCRIPTION (provided by applicant): The objective of this research is to characterize the in vitro and in vivo physiology of neurons derived from embryonic stem (ES) cells. These cells represent a potentially limitless source of pluripotent, genetically normal cells for research and therapy. Mouse ES cells can differentiate in vitro into a variety of somatic cell types including neurons, astrocytes and oligodendrocytes. In addition, differentiated ES cells survive and become morphologically integrated with surrounding host tissue following transplantation into the brain or spinal cord. Based on this work with mouse ES cells, the isolation of human ES cells has raised the possibility for novel replacement therapies in which in vitro differentiated ES cells will substitute for somatic cells lost to injury or disease. Pathologies of the nervous system that might be amenable to replacement therapy include Parkinson's disease, amyotophic lateral sclerosis, stroke, Huntington's disease, and multiple sclerosis. President Bush's decision to allow federal support for research on existing human ES lines has engendered great enthusiasm to explore the promise of stem cell-derived replacement as a new way to address these previously intractable deficits; however, much basic research remains to be done before such therapies can be achieved. Our ultimate goals are to develop procedures for efficient conversion of human ES cells into specific types of neurons and to optimize the integration of ES-derived neurons into functional networks when transplanted into a host nervous system. An essential component of functional integration is the acquisition of normal physiological properties by individual stem cell derived neurons. At this point, only limited information is available about the physiology of differentiated ES cells. Thus, the goals of this proposal are: 1) To evaluate physiological differentiation of nerve cells derived from distinct ES induction protocols. 2) To characterize the physiology of ES-derived neurons after transplantation into the brain, including rigorous tests for formation of functional synaptic connections with surrounding host neurons. Human and mouse ES cells will be used in parallel to compare their developmental potentials. All research on human ES cells will use the WA01 line in the NIH human ES cell registry.