A user-friendly scalable microfluidic platform for enhanced neuron-cell culture

用于增强神经元细胞培养的用户友好的可扩展微流体平台

• 批准号: 8660344
• 负责人: ANNE MARION TAYLOR
• 金额: $ 33.05万
• 依托单位: XONA MICROFLUIDICS, LLC
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-09 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8660344
• 关键词: Address; Adopted; Axon; Basic Science; Biochemical; Biology; Cell Culture Techniques; Cell Density; Cells; Cellular Morphology; Communities; Data; Dendrites; Development; Devices; Ensure; Extracellular Matrix Proteins; Feedback; Functional disorder; Geometry; Glass; Goals; Growth; Human; Imagery; Injury; Investigation; Label; Legal patent; Link; Lysine; Marketing; Measures; Metals; Methods; Microfluidics; Microscopy; Mind; Mission; Modeling; Modification; Molds; Nerve Tissue; Neuroglia; Neurologic; Neurons; Neurosciences; Optics; Pattern; Phase; Population; Preclinical Drug Evaluation; Preparation; Procedures; Process; Public Health; Recombinant Proteins; Research; Resolution; Rights; Sales; Scientist; Site; Surface; Synapses; Techniques; Technology; Testing; Tissue Harvesting; Toxicity Tests; Wettability; Work; base; biological research; biomaterial compatibility; commercialization; cost effective; drug discovery; human stem cells; improved; induced pluripotent stem cell; innovation; nervous system disorder; neuronal cell body; neuronal growth; novel; poly(dimethylsiloxane); presynaptic; prototype; public health relevance; synaptic function; synaptogenesis; tool; user-friendly

DESCRIPTION (provided by applicant): Neuron-cell culture is widely used for studies in basic research, drug discovery, and toxicity testing. Traditional random cultures allow limited access to subcellular compartments (axons, dendrites, synapses) due to extensive and haphazard growth of neurons. Our long-term goal is to provide robust, user-friendly, and cost effective culture platforms that can optically, fluidically, and biochemically access neurons and their subcellular compartments. Data acquired through sales of prototype platforms developed and patented by our team show a large and increasing demand. Customer feedback also indicates that technological improvements would make our platforms more accessible and user-friendly. Such improvements include ensuring greater uniformity of the device, reducing end-user assembly procedures, and enhancing viability for the long-term culturing period that is often needed for neurons. Thus, our first aim focuses on addressing these issues by (1) incorporating cell loading ports into the platform to reduce cell loading errors, (2) improving feature uniformit through the development of high-resolution, durable metal molds, (3) developing methods to reduce evaporative losses that impair neuron viability, (4) increasing the wettability and biocompatibility of the device material through surface modification, and (5) covalently linking extracellular matrix proteins onto glass to minimize end-user assembly. The successful completion of this aim will result in a consistent, cost-effective, and ready-to-use neuron-cell culture platform. Our next aim focuses on the development of novel cell-based tools to study synapses, the cornerstone of neuroscience. There is considerable demand for methods to isolate synapses and there are limitations in existing techniques. In Aim 2 we will develop user-friendly synapse isolation tools, expanding on core technological advancements from Aim 1. Aim 2 will involve (1) development of stable, artificial synaptic bead targets allowing novel investigations of the presynaptic compartment and (2) development of a three-compartment synapse isolation platform that exploits device geometry and synaptic beads to encourage synapse formation within an isolated synaptic compartment. The successful completion of this aim will result in a user-friendly, accessible, and innovative cell-based tool to optically and biochemically probe synapses. The significance of the proposed work is to improve research scientists' ability to visualize, manipulate, and measure cultured neurons leading to greater understanding of the underlying causes of neurological diseases. This research is innovative because we seek to shift current research paradigms through the development of novel cell-based tools to isolate synapses that maintain intact cell morphology in the absence of somata or glia.

描述（由申请人提供）：神经元细胞培养被广泛用于基础研究，药物发现和毒性测试的研究。传统的随机培养物可以有限地进入亚细胞室（轴突，树突，突触），这是由于神经元的广泛和随意的生长。我们的长期目标是提供强大的，用户友好且具有成本效益的文化平台，这些平台可以从光学，流动性和生化上访问神经元及其亚细胞隔室。通过我们的团队开发和专利的原型平台的销售获得的数据表明，需求不断增长。客户反馈还表明，技术改进将使我们的平台更容易访问和用户友好。这些改进包括确保设备的更大统一性，减少最终用户组装程序，并提高神经元通常需要的长期培养期的生存能力。因此，我们的第一个目的是通过（1）通过（1）将细胞加载端口纳入平台中，以减少细胞负荷误差，（2）通过开发高分辨率，耐用的金属模具来改善特征统一，（3）开发方法来减少蒸发性损失的方法，从而损害神经元的稳定性，（4）通过表达净化性和生物性的启动性（4），（4）通过净化性和生物效应（4）细胞外基质蛋白到玻璃上，以最大程度地减少最终用户组件。这个目标的成功完成将导致一个一致，成本效益且现成的神经元细胞文化平台。我们的下一个目标重点是开发基于新型细胞的工具来研究突触，即神经科学的基石。对隔离突触的方法有很大的需求，并且现有技术存在局限性。在目标2中，我们将开发对用户友好的突触隔离工具，扩展目标1的核心技术进步。目标2将涉及（1）开发稳定的人造突触珠目标，允许对突触前隔间进行新的研究，以及（2）开发三室突触隔离平台，从而开发了隔离式构造的三体突触隔离平台，以隔离式构造构造均匀的综合综合体。成功完成此目标将导致用户友好，易于访问和创新的基于细胞的工具，以光学和生化探测突触。拟议工作的重要性是提高研究科学家的可视化，操纵和测量培养的神经元的能力，从而更了解神经系统疾病的根本原因。这项研究具有创新性，因为我们试图通过开发基于细胞的新型工具来改变当前的研究范例，以隔离在没有SOMATA或GLIA的情况下保持完整细胞形态的突触。