REDOX IMAGING INDIVIDUAL EMBRYONIC STEM CELL COLONIES

单个胚胎干细胞集落的氧化还原成像

• 批准号: 8361993
• 负责人: LIN LI
• 金额: $ 1.54万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361993
• 关键词: Cell Separation; Cells; Funding; Goals; Grant; Image; Immunochemistry; Individual; Magnetic Resonance; Mediating; Microscope; Mitochondria; Mus; National Center for Research Resources; Oxidation-Reduction; Principal Investigator; Research; Research Infrastructure; Resolution; Resources; Source; Staining method; Stains; Stem cells; United States National Institutes of Health; cost; embryonic stem cell; novel; optical imaging; pluripotency; stem cell differentiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Redox state mediates embryonic stem cell (ESC) differentiation and thus offers an important complementary approach to understanding the pluripotency of stem cells. The goal of this project is to image the mitochondrial redox state of individual embryonic stem cell colonies to investigate the possible association between the redox state and the pluripotency of stem cells to ultimately develop a novel ESC sorter using the mitochondrial redox state as the cell sorting marker. The first step was set to image the redox state of the individual mouse ESC colonies without any disturbance of these colonies and to compare the result with the immunochemistry of the Oct4-stained colonies. The second step was set to transfect the ESC with another embryonic stem cell marker Nanog and simultaneously image the Nanog expression and the mitochondrial redox state. The third step was set to calibrate the redox state with appropriate agents. The fourth step was set to image the ESC colonies at the single cell resolution using a confocal microscope.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 氧化还原状态介导胚胎干细胞（ESC）分化，从而为理解干细胞的多能性提供了重要的补充方法。 该项目的目标是对单个胚胎干细胞集落的线粒体氧化还原状态进行成像，以研究氧化还原状态与干细胞多能性之间可能的关联，最终开发一种使用线粒体氧化还原状态作为细胞分选标记的新型ESC分选仪。第一步是对单个小鼠 ESC 集落的氧化还原状态进行成像，而不对这些集落进行任何干扰，并将结果与​​ Oct4 染色集落的免疫化学结果进行比较。第二步是用另一种胚胎干细胞标记 Nanog 转染 ESC，并同时对 Nanog 表达和线粒体氧化还原状态进行成像。第三步是用适当的试剂校准氧化还原态。第四步是使用共焦显微镜以单细胞分辨率对 ESC 集落进行成像。