EFRI-CBE: Regulating human embryonic stem cell differentiation via the mechanical microenvironment

PI名称：S.P.PalecekInstitution：威斯康星大学麦迪逊分校建议编号：735903EFRI：通过机械微环境调节人类胚胎干细胞分化摘要胚胎干细胞(ESCs)因其多潜能和无限增殖潜力的独特组合，在发育生物学、组织工程和再生医学应用中具有巨大的前景。要将这些细胞的可能性转化为科学和医学进步，必须了解它们如何在自我更新和分化决策中整合环境刺激，包括可溶性化学因子、细胞外基质蛋白、细胞-细胞相互作用和生物物理力。该项目将在化学定义的培养系统的背景下，具体探讨ESCs如何在空间和时间上对微环境中的机械线索做出反应。实验将确定生物物理刺激如何与生化线索协同作用，以确定胚胎干细胞是否自我更新或分化。此外，物理和化学信号影响胚胎干细胞向心肌细胞的谱系特异性分化的预测也将得到检验。要使用的方法涉及三个综合活动的迭代循环。首先，将利用首席研究人员(PI)在材料和界面方面的专业知识来设计和描述新的培养系统，这些系统允许对ESC菌落施加空间和时间上定义的机械应力。接下来，PI在细胞和分子生物学方面的经验将使人们能够阐明机械应变刺激或抑制的信号转导途径，并确定机械信号如何与ESC生长和分化的生化调节因子相互作用。最后，PI在细胞和分子建模方面的专业知识将被用来构建描述调控ESC分化的信号和路径的复杂性的数学模型。这些模型将预测微环境信号对细胞的最佳呈现，以促进自我更新或分化；然后，这些预测将得到实验验证。来自这些实验的数据将提供额外的信息，将促进进一步的模型改进。通过本项目获得的对调节ESC研究的环境信号之间关系的科学理解可能为ESC培养体系的设计提供合理的基础，从而促进其在研究和临床环境中的应用。该项目还将支持一项新的公共推广计划，包括讨论干细胞生物学和工程学的技术方面以及干细胞研究的社会影响的论坛。该项目还将推动招收高中生和本科生，特别是那些来自代表性不足群体的学生，进入干细胞工程或其他科学学科的教育倡议。最后，该项目将支持私人投资机构建设和传播干细胞工程教材。

PI name: S.P. PalecekInstitution: University of Wisconsin-MadisonProposal Number: 735903EFRI: Regulating Human Embryonic Stem Cells Differentiation via the Mechanical MicroenvironmentAbstractEmbryonic stem cells (ESCs) hold tremendous promise in developmental biology, tissue engineering, and regenerative medicine applications because of their unique combination of pluripotency and limitless proliferation potential. To translate the possibilities of these cells into scientific and medical advances, one must understand how they integrate environmental stimuli, including soluble chemical factors, extracellular matrix proteins, cell-cell interactions, and biophysical forces, in their self-renewal and differentiation decisions. This project will specifically address how ESCs spatially and temporally respond to mechanical cues in the microenvironment in the context of a chemically defined culture system. Experiments will ascertain how biophysical stimuli synergize with biochemical cues to determine whether an ESC self-renews or differentiates. In addition, the predication that both physical and chemical signals influence lineage-specific differentiation of ESCs to cardiac myocytes will be tested. The approach to be used involves an iterative loop of three integrated activities. First, the Principal Investigators' (PIs') expertise in materials and interfaces will be used to design and characterize novel culture systems that permit application of spatially and temporally defined mechanical stresses to ESC colonies. Next, the PIs' experience in cell and molecular biology will allow one to elucidate signal transduction pathways stimulated or repressed by mechanical strain and to determine how mechanical signals interact with biochemical regulators of ESC growth and differentiation. Finally, the PIs' expertise in cellular and molecular modeling will be used to construct mathematical models describing the complexity of signals and pathways regulating ESC differentiation. The models will predict optimal presentation of microenvironmental cues to cells to promote self-renewal or differentiation; then, these predictions will be tested experimentally. The data from these experiments will provide additional information that will promote further model refinement. The scientific understanding of the relationships between environmental signals regulating ESC research gained from this project may provide a rational basis for design of ESC culture systems that will facilitate their use in research and clinical settings. The project will also support a novel public outreach plan, consisting of forums discussing the technical aspects of stem cell biology and engineering as well as societal impacts of ESC research. Educational initiatives to recruit high school and undergraduate students, especially those from underrepresented groups, into stem cell engineering or other scientific disciplines will also be enabled by this project. Finally, this project will support construction and dissemination of stem cell engineering educational materials by the PIs.