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.

流式细胞仪和细胞分类核心（核心D）的功能是为每个项目提供流式细胞仪分析所需的专业知识，设备和设施以及心脏祖细胞研究所需的细胞分类程序（CPC）。在复杂的异质种群中，准确的鉴定，表征和定量在技术上是要求的，需要专业且昂贵的设备和人员。该程序的MAIOR优势之一是，它将为所有四个方面提供流式细胞仪和细胞分类的设施和技术。 否则，任何孤立的竞争都无法负担得起。基于流式细胞仪的分析，用于鉴定特定细胞群体，细胞承诺或表型的变化以及细胞功能（例如细胞周期，生存力，信号转导或电位）的表征对所有项目至关重要。因此 实现计划项目的目标。除了效率和成本节省的明显优势外，核心还将使四名具有多样化和互补背景的研究人员（RokoshDr。Rokosh，McCracken，Ratajczak和O'Toole）能够结合其技术和生物学专业知识，以确保已成功执行所提出的工作。 核心将为计划项目调查人员提供识别和表征复杂种群特定细胞类型的能力。具体而言，核心将对各种细胞的标记表达进行分析，包括CPC和更多分化的细胞，包括分化过程中标记表达的变化。检测自由基的细胞内产生（即，将DAFDA用于NO，而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； 确定细胞信号的变化；利用多个标记物分离干细胞；并在细胞周期的每个阶段量化细胞。核心还积极地识别和表征对各种项目有用的新型细胞标记。