Subcellular Targeting of Dosant Streams in Culture

培养中剂量流的亚细胞靶向

• 批准号: 7132387
• 负责人: JASON Ben SHEAR
• 金额: $ 16.15万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7132387
• 关键词: biotechnology; cell line; chemical structure function; experimental designs; fluid flow; lasers; membrane model; model design /development; neurons; physical chemical interaction; technology /technique development; time resolved data; tissue /cell culture

DESCRIPTION (provided by applicant): Two-dimensional (2D) cultures provide important model systems for studying properties of neurons and other cells in relatively simple and controllable environments. Key to value of this reductionist approach is development of technologies for perturbing and characterizing neuronal properties at the subcellular, single-cell, and cellular-ensemble levels. Although valuable methods exist for interfacing with cells electronically, current techniques for interacting chemically with neurons provide inadequate spatial control, particularly in applications that require dosings to be sustained for periods of seconds or longer. The overall objective of this project is to develop a generalized approach for chemically interfacing with neuronal cultures at numerous sites in parallel and with arbitrary, real-time control over interaction coordinates. The methods developed will be applicable to a broad range of cell biology problems in neuroscience and beyond, including chemotaxis and stem-cell differentiation. Specifically, the project is focused on development of an approach for rapid, laser-mediated introduction of apertures within membranes that serve as supports for cell growth as well as separating two flow chambers: a cell-medium chamber and an underlying dosant chamber. Aperture creation, caused by point ablation of the thin membrane barrier, introduces a laminar-flow stream from the higher pressure dosant chamber into the cell environment. Specific Aim 1 is directed toward the development of a reproducible and well-characterized platform, with a focus on evaluation of membrane materials and characterization of spatiotemporal limits of the system. Aim 2 focuses on expanding the functionality of the platform through the implementation of reversible valving of apertures and a means to modify flow-stream directionality. Aim 3 is directed toward initial demonstration of the utility of this tool for stimulating and modulating cell-line and neuronal cultures at multiple points-of-interest in parallel.

描述（由申请人提供）：二维（2D）培养物为在相对简单且可控的环境中研究神经元和其他细胞的特性提供了重要的模型系统。这种还原论方法的价值关键是开发在亚细胞、单细胞和细胞整体水平上扰乱和表征神经元特性的技术。尽管存在与细胞电子连接的有价值的方法，但当前与神经元化学相互作用的技术提供了不足的空间控制，特别是在需要持续给药数秒或更长时间的应用中。该项目的总体目标是开发一种通用方法，用于与多个位点的神经元培养物并行化学连接，并对相互作用坐标进行任意、实时控制。开发的方法将适用于神经科学及其他领域的广泛细胞生物学问题，包括趋化性和干细胞分化。具体来说，该项目的重点是开发一种通过激光介导在膜内快速引入孔径的方法，作为细胞生长的支持以及分隔两个流动室：细胞介质室和下面的剂量室。由薄膜屏障的点烧蚀引起的孔径形成，将层流从高压剂量室引入到细胞环境中。具体目标 1 旨在开发可重复且特征良好的平台，重点是膜材料的评估和系统时空限制的表征。目标 2 侧重于通过实施孔径可逆阀门和修改流方向性的方法来扩展平台的功能。目标 3 旨在初步演示该工具在多个兴趣点并行刺激和调节细胞系和神经元培养物的实用性。