权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Cell-based sensors for measuring impact of microsystems on cell physiology

基于细胞的传感器，用于测量微系统对细胞生理学的影响

基本信息

批准号：
8409794
负责人：
Joel Voldman
金额：
$ 32.32万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-01 至 2014-12-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): There is a steady increase in the development of microsystems for manipulating, measuring, culturing, and separating cells. Important to the design and adoption of these systems is knowledge of how the systems themselves alter cell state. Such measurements can be difficult to perform, due to either lack of sophistication on the part of the user, or difficulty performing conventional assays with limited numbers of cells in small devices. As a result, assays to measure the impact of microsystems on cell physiology are typically limited to general characteristics such as viability, morphology, and growth. This proposal focuses on developing a suite of cell lines that will provide a more nuanced view of the effects of stress on cell physiology. Building on prior work in our lab creating a heat shock cell sensor, we propose to create a set of cell lines that report on transcriptional activation of stress response pathways relevant to cell-based microfluidic devices. Specifically, we believe that lines reporting on DNA damage, shear stress, and heat shock will provide a valuable set of reagents for designers & users of microsystems. We propose to create these cell lines so that they are spectrally distinct and thus can be mixed and assayed at once. We will formulate a set of standards for applying these reagents and interpreting results from them. As such, we propose to not only generate the cell lines, but the requisite protocols, imaging and data interpretation algorithms, as well as case studies for others to follow. Our specific aims are to (1) create cell lines with inducible fluorescent proteins under the control heat shock-, DNA damage-, and shear stress-inducible promoters. Each cell line will be created in three cell types, representing the diversity of cells one may use in a microsystem; (2) Determine the responses of the cell lines to input stresses, such as shear, heat, and light; (3) Undertake a case study of the use of these reagents and distribute them to the community.

描述(由申请人提供)：用于操纵、测量、培养和分离细胞的微系统的发展正在稳步增加。对于这些系统的设计和采用来说，重要的是了解这些系统本身如何改变细胞状态。这样的测量可能很难进行，这要么是因为用户缺乏复杂性，要么是因为在小型设备中使用有限数量的细胞进行传统的分析是困难的。因此，衡量微系统对细胞生理学影响的分析通常仅限于一般特征，如活力、形态和生长。这项建议的重点是开发一套细胞系，提供对压力对细胞生理学影响的更微妙的看法。在我们实验室创建热休克细胞传感器的先前工作的基础上，我们建议创建一组细胞系，报告与基于细胞的微流控设备相关的应激反应通路的转录激活。具体地说，我们相信，报告DNA损伤、剪切力和热震的LINES将为微系统的设计者和用户提供一套有价值的试剂。我们建议创建这些细胞系，以便它们在光谱上是不同的，从而可以一次混合并进行分析。我们将制定一套使用这些试剂的标准，并解释它们的结果。因此，我们建议不仅要生成细胞系，还要生成必要的协议、成像和数据解释算法，以及供其他人效仿的案例研究。我们的具体目标是(1)在热休克、DNA损伤和剪切力诱导启动子的控制下，建立具有诱导荧光蛋白的细胞系。每个细胞系将被创建为三种细胞类型，代表人们可以在微系统中使用的细胞的多样性；(2)确定细胞系对输入应力(如剪切、热和光)的反应；(3)进行这些试剂的使用案例研究，并将其分发给社区。