Multi-Well Extracellular Flux Analyzer

描述(申请人提供)：在动物细胞中有两种途径产生大量的三磷酸腺苷-线粒体氧化磷酸化和糖酵解。生物学家对这些ATP产生过程的调控非常感兴趣，但在培养细胞中很难研究这些过程，特别是当细胞以多孔配置培养以获得高产量时。最近，海马生物科学公司开发了XF24胞外通量分析仪，可以连续测量在24孔板中培养的细胞的O2消耗、酸形成和CO2形成。通量分析仪测量的三个参数允许研究人员重复和非破坏性地测量单细胞培养中的氧化磷酸化、糖酵解、营养利用和其他过程。因此，我们的具体目标是1)获得一台用于测量氧气摄取、酸释放和二氧化碳产生的海马三分析仪胞外通量分析仪，以及2)将该仪器置于多用户环境中，为个别用户项目提供技术支持、用户培训和应用专业知识。这项新兴技术将独特地允许NIH资助的研究人员从事各种项目，以评估正常生理和疾病模型中的生物能量学和代谢反应，包括癌症、酒精性肝毒性、黄斑变性、心脏缺血和衰竭、多器官衰竭和糖尿病。这些研究的结果可能会导致为其中一些疾病开发新的治疗方法。

DESCRIPTION (provided by applicant): Two pathways generate the bulk of ATP in animal cells - mitochondrial oxidative phosphorylation and glycolysis. Regulation of these ATP-generating processes is of fundamental interest to biologists, but these processes are difficult to study in cultured cells, especially when the cells are cultured in a multiwell configuration for high throughput. Recently, Seahorse Bioscience has developed the XF24 Extracellular Flux Analyzer that permits serial measurements of O2 consumption, acid formation and CO2 formation of cells cultured in 24- well plates. The three parameters measured by the flux analyzer permit investigators to measure repetitively and non-destructively oxidative phosphorylation, glycolysis, nutrient utilization and other processes in single cell cultures. Accordingly, our specific aims are to 1) acquire a Seahorse Three-Analyte Extracellular Flux Analyzer for measurement of O2 uptake, acid release and CO2 generation, and 2) place this instrument in a multi-user environment that provides technical support, user training and applications expertise in support of individual user projects. This emerging technology will uniquely permit NIH-funded researchers working on a variety of projects to assess bioenergetics and metabolic responses in normal physiology and models of disease, including cancer, alcoholic hepatotoxicity, macular degeneration, heart ischemia and failure, multiple organ failure and diabetes. The results of those studies could lead to the development of new therapeutic treatments for some of these diseases.