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单元发送指令，这些信号允许运动，感觉，记忆，学习以及无数在日常生活中授予的其他活动。神经细胞功能的丧失导致创伤性损伤，例如枪伤或汽车事故以及神经退行性疾病。这种健康问题的规模几乎是不可估量的：仅400万美国人被诊断出患有阿尔茨海默氏症，另外500,000例患有帕金森氏病。大多数神经元是由神经干细胞或神经元基因细胞在胚胎发育过程中产生的。生物医学研究人员从人类胚胎干细胞（HESC）衍生了神经元素细胞，通过体外分化提供了人类神经元和神经胶质细胞的潜在来源。现有的人类神经源性源的来源被传播为神经球的悬浮液，这些悬浮液无法轻易适应定量分析或用于治疗化合物或神经毒性测试的高通量或高含量筛选。由于缺乏足够的人类神经细胞模型，为减轻神经系统疾病负担而减轻神经系统疾病负担的生物医学研究受到了阻碍。 Aruna生物医学建议开发包含所需试剂的转换式套件，以将WA09 HESC（NIH注册）衍生成神经元素构成神经元和神经胶质细胞的原代培养物。潜在的技术创新是在粘附的单层培养物中增殖和区分Aruna神经源性的能力。预期的结果是，研究人员将增加获得定量研究的机会。在这个快速轨道STTR项目的第一阶段部分中，人类神经元素构成将源自WA09 hESC。免疫化学方法和流动细胞细胞仪将验证扩展到纯度为90％并保留正常二倍体核型的培养物。为了证明概念证明，将在体外区分神经元素，并且将通过共聚焦免疫荧光显微镜确定分化细胞培养物中运动神经元的产率，并在三个试验中进行了量化，并使用统计方法进行了测试。 I期研究的结果将为快速轨道II期提案提供基础，以优化商业生产方法，并通过最终用户增强神经元素细胞及其衍生物的实用性。 Aruna生物医学的愿景是，我们在第三阶段对产品的商业化将通过增强学术和商业研究人员使用基于hESC的神经细胞基于基于神经细胞的测定的能力来大大推动这一领域。

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.