Neural cell based assays derived from human ES cells

基于人类 ES 细胞的神经细胞检测

• 批准号: 6998993
• 负责人: STEVEN L STICE
• 金额: $ 29.37万
• 依托单位: ARUNA BIOMEDICAL, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6998993
• 关键词: cell differentiation; cell proliferation; confocal scanning microscopy; cryopreservation; fibroblasts; flow cytometry; glia; human embryonic stem cell line; immunofluorescence technique; karyotype; motor neurons; nerve stem cell; neurons; statistics /biometry; technology /technique development; tissue /cell culture

DESCRIPTION (provided by applicant): Neurons send instructions to other cells in the form of electrical signals that allow movement, sensation, memory, learning and countless other activities that are taken for granted in daily life. Loss of neural cell function results from traumatic injuries such as gunshots wounds or car accidents and from neurodegenerative diseases. The scale of this health problem is virtually immeasurable: 4 million Americans alone are now diagnosed with Alzheimer's and another 500,000 with Parkinson's disease. Most neurons are produced during embryonic development by neural stem cells or neuroprogenitor cells. Biomedical researchers have derived neuroprogenitor cells from human embryonic stem cells (hESCs), providing a potential source of human neurons and glial cells for biomedical research through in vitro differentiation. Existing sources of human neuroprogenitors are propagated as suspensions of neurospheres that cannot be easily adapted for quantitative analysis or use in high-throughput or high-content screens for therapeutic compounds or for tests of neurotoxicity. Biomedical research to relieve the burden of neurological disorders is hampered by the lack of an adequate human neural cell-based model. Aruna Biomedical proposes to develop turn-key kits containing the needed reagents to propagate and reliably differentiate WA09 hESC (NIH registered) derived neuroprogenitors into primary cultures of neurons and glial cells. The potential technical innovation is the ability to proliferate and differentiate Aruna's neuroprogenitors in adherent monolayer cultures. The expected outcome is that researchers will have increased access to quantitative studies. In the Phase I portion of this Fast Track STTR project, human neuroprogenitors will be derived from WA09 hESCs. Immunochemical methods and flow cell cytometry will verify expansion into cultures that are s 90% pure and retain a normal diploid karyotype. To demonstrate proof of concept, neuroprogenitors will be differentiated in vitro and the yield of motor neurons in differentiated cell cultures will be established by confocal immunofluorescence microscopy, quantified in three trials, and tested with statistical methods. The results of Phase I studies will provide the groundwork for a Fast Track Phase II proposal to optimize methods for commercial production and to enhance the utility of neuroprogenitor cells and their derivatives by the end user. The vision of Aruna Biomedical is that commercialization of our products in Phase III will significantly advance the field by enhancing the ability of academic and commercial researchers to perform quantitative analysis with hESC derived neural cell-based assays.

描述（由申请人提供）：神经元以电信号的形式向其他细胞发送指令，使运动、感觉、记忆、学习和无数其他日常生活中理所当然的活动成为可能。神经细胞功能的丧失是由创伤性损伤，如枪伤或车祸以及神经退行性疾病造成的。这一健康问题的规模实际上是无法估量的：目前仅美国就有400万人被诊断患有阿尔茨海默氏症，另有50万人患有帕金森病。大多数神经元是在胚胎发育期间由神经干细胞或神经祖细胞产生的。生物医学研究人员已经从人类胚胎干细胞（hESCs）中获得了神经祖细胞，通过体外分化为生物医学研究提供了人类神经元和神经胶质细胞的潜在来源。现有的人类神经祖细胞来源以神经球悬浮液的形式繁殖，不容易用于定量分析或用于治疗性化合物的高通量或高含量筛选或用于神经毒性测试。由于缺乏适当的基于人类神经细胞的模型，减轻神经疾病负担的生物医学研究受到阻碍。Aruna Biomedical提议开发包含所需试剂的交钥匙试剂盒，以繁殖和可靠地将WA09 hESC （NIH注册）衍生的神经祖细胞分化为神经元和胶质细胞的原代培养。潜在的技术创新是在贴壁单层培养中增殖和分化阿鲁纳神经祖细胞的能力。预期的结果是，研究人员将有更多的机会进行定量研究。在这个快速通道STTR项目的I期部分，人类神经祖细胞将来自WA09 hESCs。免疫化学方法和流式细胞术将证实培养物的扩增纯度为90%，并保留正常的二倍体核型。为了证明这一概念，神经祖细胞将在体外分化，分化细胞培养中的运动神经元的产量将通过共聚焦免疫荧光显微镜确定，在三个试验中量化，并用统计方法进行测试。I期研究的结果将为快速通道II期提案提供基础，以优化商业化生产的方法，并提高最终用户对神经祖细胞及其衍生物的效用。Aruna Biomedical的愿景是，通过提高学术和商业研究人员使用hESC衍生神经细胞进行定量分析的能力，我们的产品在III期的商业化将显著推进该领域的发展。

项目成果

Human neural progenitors express functional lysophospholipid receptors that regulate cell growth and morphology.

• DOI: 10.1186/1471-2202-9-118
• 发表时间: 2008-12-11
• 期刊: BMC neuroscience
• 影响因子: 2.4
• 作者: Hurst JH;Mumaw J;Machacek DW;Sturkie C;Callihan P;Stice SL;Hooks SB
• 通讯作者: Hooks SB

Ion channels and ionotropic receptors in human embryonic stem cell derived neural progenitors.

• DOI: 10.1016/j.neuroscience.2011.04.039
• 发表时间: 2011-09-29
• 期刊: NEUROSCIENCE
• 影响因子: 3.3
• 作者: Young, A.;Machacek, D. W.;Dhara, S. K.;MacLeish, P. R.;Benveniste, M.;Dodla, M. C.;Sturkie, C. D.;Stice, S. L.
• 通讯作者: Stice, S. L.