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

• 批准号: 8010388
• 负责人: Joel Voldman
• 金额: $ 33.49万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8010388
• 关键词: Adoption; Affect; Algorithms; Biological Assay; Biotechnology; Case Study; Cell Count; Cell Line; Cell Separation; Cell physiology; Cells; Characteristics; Collecting Cell; Color; Communities; Cyan Fluorescent Protein; DNA Damage; Data; Development; Devices; Ensure; Growth; HSF1; Heat-Shock Response; Heating; Image; Image Analysis; Knowledge; Life; Light; MCF7 cell; Measurement; Measures; Metric; Microfluidic Microchips; Morphology; Pathway interactions; Proteins; Protocols documentation; Reagent; Reporter; Reporting; Staining method; Stains; Stress; System; Techniques; Testing; Transcriptional Activation; Work; base; biological adaptation to stress; cell type; design; electric field; experience; microsystems; point-of-care diagnostics; promoter; public health relevance; response; sensor; shear stress

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. PUBLIC HEALTH RELEVANCE: Microsystems that can analyze small numbers of cells could have wide use for point-of- care diagnostics and biotechnology. Currently, there are no standardized metrics by which such microsystems can be designed or used to ensure that they are not harmful to the cells they are trying to analyze. We are proposing to develop cell "sensors" that would glow colors if they are subjected to stresses from microsystems.

描述（由申请人提供）：用于操纵，测量，培养和分离细胞的微生系统的发展稳定增加。对于这些系统的设计和采用至关重要的是了解系统本身如何改变细胞状态的知识。由于用户缺乏复杂性，或者难以执行小型设备中数量有限的传统测定，因此很难执行此类测量。结果，测量微生系统对细胞生理的影响的测定通常仅限于一般特征，例如生存力，形态和生长。该提案着重于开发一系列细胞系，这些细胞系将对压力对细胞生理的影响提供更细微的看法。在我们实验室的先前工作的基础上，我们建议创建一组细胞系，以报告与基于细胞的微流体设备相关的应力响应途径的转录激活。具体来说，我们认为报告有关DNA损伤，剪切应力和热量冲击的线路将为微型系统的设计师和使用者提供一套宝贵的试剂。我们建议创建这些细胞系，以使它们在频谱上不同，因此可以立即混合和测定。我们将制定一组应用这些试剂并解释其结果的标准。因此，我们建议不仅生成细胞系，还提出必要的协议，成像和数据解释算法以及案例研究，以供其他人遵循。我们的具体目的是（1）在对照热冲击，DNA损伤和剪切应力诱导启动子下创建具有诱导荧光蛋白的细胞系。每个细胞系将在三种细胞类型中创建，代表一个人可能在微系统中使用的细胞多样性。 （2）确定细胞系对输入应力的响应，例如剪切，热和光； （3）对这些试剂的使用进行案例研究，并将其分配给社区。 公共卫生相关性：可以分析少量细胞的微型系统可以广泛用于医疗点诊断和生物技术。当前，没有标准化指标可以设计或使用此类微型系统来确保它们对试图分析的单元不构成危害。我们提议开发细胞“传感器”，如果它们受到微系统的应力，它们会发光颜色。