STEM CELL-MYOCYTE ELECTRICAL COUPLING VIA A LASER PATTERNED CELL BRIDGE

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 我们的长期目标是建立一个具有特定微环境的细胞共培养模型，在单细胞水平上系统地研究成体干细胞与心肌细胞的机电耦合。本项目的目的是了解骨髓干细胞如何与心肌细胞电偶联，以及干细胞分化将如何影响这种偶联。我们将研究骨髓干细胞在工程心肌细胞共培养模型中的电偶联。在该模型中，将制造相同的细胞培养微孔，以允许通过应用激光束在每个孔中创建限定的干细胞-肌细胞界面。 为了检验干细胞和肌细胞之间的电耦合将受细胞的空间排列和干细胞分化的时间阶段调节的假设，将解决以下具体目标： * 在细胞水平上确定干细胞和共培养的心肌细胞之间电耦合的时间和空间范围 * 目的：研究不同连接蛋白在干细胞-心肌细胞电偶联中的作用 * 研究干细胞分化对干细胞与心肌细胞电偶联的影响 心脏病是美国男性和女性的主要死因，占每年死亡人数的近40%。通过增加骨髓干细胞和心肌细胞之间电耦合的现有知识，我们的研究将有助于再生医学最近的努力，利用患者自身的细胞恢复受损心脏的功能。这项研究的意义在于它提供了一个实验模型，通过该模型，各种干细胞在其功能分化期间受到高度控制的微环境。在此模型中应用激光技术，可以研究干细胞和肌细胞之间的电耦合和整合的确切时间和空间约束。这项研究的结果将显着推进我们的干细胞和肌细胞之间的功能整合的基础知识。 除了50%的释放时间，克莱姆森将提供超过2,500平方英尺的实验室空间和无限制地访问所有核心设施，这是工作人员和提供最先进的设备。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our long-term goal is to establish a cell coculture model with a defined microenvironment to systematically study the electromechanical coupling of adult stem cells with cardiac myocytes at the single-cell level. The objective for this project is to understand how bone marrow stem cells electrically couple with cardiac myocytes and how stem cell differentiation will affect this coupling. We will investigate the electrical coupling of bone marrow stem cells in an engineered cardiac myocyte-coculture model. In this model, identical cell-culture microwells will be fabricated to allow creation of a defined stem cell-myocyte interface in each well through application of a laser beam. To test the hypothesis that the electrical coupling between stem cells and myocytes will be regulated by the spatial arrangements of cells and the temporal stages of stem cell differentiation, the following specific aims will be addressed: * To determine temporal and spatial extent of the electrical coupling between stem cells and cocultured myocytes at the cellular level * To identify effects of different connexins in stem cell-myocyte electrical coupling * To determine teffects of stem cell differentiation on stem cells' electrical coupling with cardiac myocytes Heart disease is America's leading cause of death for both men and women, accounting for nearly 40% of all annual deaths. By adding to the current knowledge of electrical coupling between bone marrow stem cells and heart muscle cells, our research will contribute to the recent effort in regenerative medicine to restore the function of a damaged heart using the patient's own cells. The significance of this research is that it provides an experimental model by which various stem cells are subjected to a highly controlled microenvironment during their functional differentiation. With application of the laser technique in this model, exact temporal and spatial constraints on electrical coupling and integration between stem cells and myocytes can be investigated. The results of this research will significantly advance our fundamental knowledge of the functional integration between stem cells and myocytes. In addition to 50% release time, Clemson will provide more than 2,500 sq ft of lab space and unlimited access to all Core facilities, which are staffed and provide with state-of-the-art equipment.