Microchip-based Cell Reactor Analysis System

基于微芯片的电池反应器分析系统

• 批准号: 8100044
• 负责人: ROBERT Scott MARTIN
• 金额: $ 28.5万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-10 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100044
• 关键词: Biological Process; Brain; Cardiovascular system; Catecholamines; Cell Communication; Cells; Chronic; Cystic Fibrosis; Detection; Development; Devices; Disease; Dopamine; Electrochemistry; Electrophoresis; Endothelial Cells; Endothelium; Erythrocytes; Goals; Grant; Hypoxia; Immobilization; Immobilized Cells; Impairment; Implant; Inflammation; Laboratories; Lead; Microchip Electrophoresis; Microdialysis; Microfluidic Microchips; Microglia; Molecular; Monitor; Muscle relaxation phase; Neuroglia; Neurons; Neurotransmitters; Nitric Oxide; Norepinephrine; Onset of illness; Parkinson Disease; Performance; Play; Process; Production; Pulmonary Hypertension; Reactive Oxygen Species; Reporting; Research Personnel; Resolution; Role; Sampling; Smooth Muscle; Stream; Substantia nigra structure; System; Systems Analysis; Technology; Translating; Vasodilation; base; cell type; dopaminergic neuron; in vitro Model; in vivo; interest; micro-total analysis system; microchip

DESCRIPTION (provided by applicant): The overall goal of this AREA renewal application is to develop microchip approaches that can be used to study cell-to-cell interactions at the molecular level. There are many examples in which the interaction of different cell types plays a role in normal biological function or in the onset of disease. One example is the interaction between neurons and glia cells that have undergone inflammation. It has been suggested that the degeneration of dopaminergic neurons that is prevalent in Parkinson's disease may be related to chronic inflammation of microglia, with the microglia producing nitric oxide and other reactive oxygen species that interact with the neurons. Another example is the vasodilatation process, where it has been shown that red blood cells, when exposed to hypoxic conditions or deformation, interact with endothelial cells via ATP release, leading to the production of nitric oxide and subsequent smooth muscle relaxation. Impairment of the ATP release from red blood cells leads to less nitric oxide production and has been postulated to play a role in several diseases. While there have been numerous examples of immobilizing cells on-chip and detecting a specific analyte released from the cells, there have been few reports of integrating multiple cell types on chip where cell-to-cell communication can be studied in a manner where numerous neurotransmitters/products can be monitored. In this grant we propose to continue the development of microchip approaches that enable the integration of cell immobilization with a general analysis step (electrophoresis), where a variety of analyses that are either released or transported through a layer of cells can be separated and subsequently detected via electrochemistry. Importantly, we propose to develop technology that will enable a researcher to monitor cell- to-cell communication between 2 layers of immobilized cells (PC 12 cells and microglia cells) or between a flowing stream of cells (red blood cells) and a layer of immobilized cells (endothelial cells). In addition, we propose to expand this microchip approach to enable in vivo studies by integrating microdialysis sampling, which can be used to study the interactions of different cell types by stereotaxically implanting a probe in the region of interest, with segmented flow, microchip electrophoresis, and electrochemical detection. The specific aims of the grant are to 1) Use of reservoir-based cell immobilization and microchip electrophoresis with electrochemical detection to study the interaction between multiple cell types; 2) Develop a microchip device that can be used to study cell-to-cell communication between red blood cells and an immobilized endothelium; and 3) Integration of microdialysis sampling and segmented flow with microchip electrophoresis and electrochemical detection for in vivo sampling. PUBLIC HEALTH RELEVANCE: The overall goal of this AREA renewal application is to develop microchip approaches that can be used to study cell-to-cell interactions at the molecular level. The resulting technology will enable monitoring of cell-to- cell communication between 2 layers of immobilized cells or between flowing cells and an immobilized endothelium. The approaches will also enable in vivo studies by integrating microdialysis sampling with segmented flow, microchip electrophoresis, and electrochemical detection.

描述（由申请人提供）：本AREA更新申请的总体目标是开发可用于在分子水平上研究细胞间相互作用的微芯片方法。在许多例子中，不同细胞类型的相互作用在正常的生物功能或疾病的发病中起作用。一个例子是经历炎症的神经元和神经胶质细胞之间的相互作用。有研究表明，帕金森病中普遍存在的多巴胺能神经元的退化可能与小胶质细胞的慢性炎症有关，小胶质细胞产生一氧化氮和其他活性氧与神经元相互作用。另一个例子是血管舒张过程，研究表明，当红细胞暴露于缺氧条件或变形时，通过ATP释放与内皮细胞相互作用，导致一氧化氮的产生和随后的平滑肌松弛。红细胞ATP释放的损伤导致一氧化氮产生减少，并被认为在几种疾病中起作用。虽然已经有许多将细胞固定在芯片上并检测从细胞中释放的特定分析物的例子，但很少有将多种细胞类型整合到芯片上的报道，在芯片上可以以一种可以监测许多神经递质/产物的方式研究细胞间的通信。在这项资助中，我们建议继续开发微芯片方法，使细胞固定与一般分析步骤（电泳）相结合，在电泳中，通过细胞层释放或运输的各种分析可以被分离并随后通过电化学检测。重要的是，我们建议开发一种技术，使研究人员能够监测两层固定细胞（pc12细胞和小胶质细胞）之间或细胞流（红细胞）和固定细胞层（内皮细胞）之间的细胞间通信。此外，我们建议通过整合微透析采样来扩展这种微芯片方法，通过在感兴趣的区域立体定向植入探针，利用分段流，微芯片电泳和电化学检测来研究不同细胞类型的相互作用，从而实现体内研究。拨款的具体目的是：1)利用基于水库的细胞固定化和微芯片电泳与电化学检测来研究多种细胞类型之间的相互作用；2)开发可用于研究红细胞与固定内皮细胞间细胞间通讯的微芯片装置；3)将微透析取样和分段流与微芯片电泳和电化学检测相结合，用于体内取样。