Flow Cytometry and Cell Sorting Core

流式细胞术和细胞分选核心

• 批准号: 8188515
• 负责人: GREGG ROKOSH
• 金额: $ 22.33万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8188515
• 关键词: Area; Biological; Budgets; Cardiac; Cardiomyopathies; Cell Cycle; Cell Separation; Cell Survival; Cell physiology; Cells; Centers of Research Excellence; Complex; Cost Savings; Coupled; Diabetes Mellitus; Ensure; Equipment; Flow Cytometry; Foundations; Funding; Generations; Goals; Human Resources; Institutes; Maintenance; Membrane Potentials; Myocardial Ischemia; Nature; Obesity; Phase; Phenotype; Population; Population Heterogeneity; Procedures; Reperfusion Therapy; Research Personnel; Signal Transduction; Stem cells; Techniques; United States National Institutes of Health; Work; base; cell type; diacetyldichlorofluorescein; mitochondrial membrane; novel; operation; programs; stem cell biology; stem cell therapy

The function of the Flow Cytometry and Cell Sorting Core (Core D) is to provide each Project with the expertise, equipment, and facilities required for the flow cytometric analyses and cell sorting procedures that are needed for studies of cardiac progenitor cells (CPCs). Accurate identification, characterization, and quantitation of cells in a complex heterogeneous population are technically demanding and require specialized and expensive equipment and personnel. One of the maior advantages of this Program Proiect is that it will make available to all four Proiects the facilities and techniques of flow cytometry and cell sorting. which otherwise would not be affordable to any Proiect in isolation. Flow cytometry-based analysis for the identification of specific cell populations, changes in cell commitment or phenotype, and characterization of cell functions such as cell cycle, viability, signal transduction, or potential, will be crucial to all Projects. A central common facility that performs these technically challenging analyses is therefore indispensable to achieve the goals of the Program Project. Besides the obvious advantages of efficiency and cost savings, the Core will also enable four investigators with diverse and complementary backgrounds (Drs. Rokosh, McCracken, Ratajczak, and O'Toole) to combine their technical and biological expertise in order to ensure that the proposed work is successfully executed. The Core will offer the Program Project investigators the ability to identify and characterize specific cell types from complex populations. Specifically, the Core will perform analysis of marker expression in various cells, including CPCs and more differentiated cells and including changes in marker expression during differentiation; detect intracellular generation of radicals (i.e., using DAFDA for NO and DCFDA for ROS); examine mitochondrial membrane integrity, membrane potential, and function; assess cell viability; determine changes in cell signaling; isolate stem cells utilizing multiple markers; and quantify cells at each phase of the cell cycle. The Core is also active in identifying and characterizing novel cell markers that will be useful for the various Projects.

流式细胞术和细胞分选核心（核心D）的功能是为每个项目提供心脏祖细胞（CPC）研究所需的流式细胞术分析和细胞分选程序所需的专业知识、设备和设施。在复杂的异质群体中准确鉴定、表征和定量细胞在技术上要求很高，并且需要专门且昂贵的设备和人员。该项目的主要优势之一是它将为所有四个项目提供流式细胞术和细胞分选的设备和技术。 否则任何孤立的项目都负担不起。基于流式细胞术的分析，用于识别特定细胞群、细胞定型或表型的变化以及细胞功能（如细胞周期、活力、信号转导或潜能）的表征，对所有项目都至关重要。因此，一个执行这些技术上具有挑战性的分析的中央共用设施是必不可少的， 实现计划项目的目标。除了效率和成本节约的明显优势外，核心还将使四名具有不同和互补背景的研究者（Rokosh、麦克拉肯、Ratajczak和O 'Toole博士）能够联合收割机结合他们的技术和生物学专业知识，以确保拟议的工作成功执行。 核心将为计划项目研究人员提供从复杂群体中识别和表征特定细胞类型的能力。具体地，核心将对各种细胞中的标志物表达进行分析，包括CPC和更分化的细胞，并包括分化期间标志物表达的变化;检测细胞内自由基的产生（即，使用DAFDA用于NO和DCFDA用于ROS）;检查线粒体膜完整性、膜电位和功能;评估细胞活力; 确定细胞信号传导的变化;利用多种标记物分离干细胞;以及在细胞周期的每个阶段对细胞进行定量。核心还积极识别和表征新的细胞标记物，这将有助于各种项目。