Physiology of Neurons from Human & Mouse ES Cells

描述（由申请人提供）：本研究的目的是表征胚胎干（ES）细胞衍生的神经元的体外和体内生理学。这些细胞代表了用于研究和治疗的多能、遗传正常细胞的潜在无限来源。小鼠ES细胞在体外可分化为神经元、星形胶质细胞和少突胶质细胞等多种体细胞类型。此外，分化的ES细胞存活，并在移植到脑或脊髓后与周围宿主组织形成形态学整合。基于小鼠ES细胞的这项工作，人ES细胞的分离提高了新的替代疗法的可能性，其中体外分化的ES细胞将替代因损伤或疾病而丧失的体细胞。可能适合替代疗法的神经系统病理包括帕金森病、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支持现有的人类胚胎干细胞系的研究，这引起了人们极大的热情，以探索干细胞衍生替代作为解决这些以前难以解决的缺陷的新方法的前景;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程序，有效地将人类ES细胞转化为特定类型的神经元，并优化ES衍生的神经元移植到宿主神经系统时整合到功能网络中。功能整合的一个重要组成部分是个体干细胞衍生的神经元获得正常的生理特性。在这一点上，只有有限的信息是关于分化的ES细胞的生理学。因此，本建议的目标是：1）评估来自不同ES诱导方案的神经细胞的生理分化。2)表征ES衍生神经元移植入脑后的生理学，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ES细胞将平行使用，以比较它们的发育潜力。所有关于人类ES细胞的研究都将使用NIH人类ES细胞登记处的WA 01细胞系。

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.