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

PI姓名：S. P. Palecek机构：威斯康星大学麦迪逊分校提案编号：735903 EFRI：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）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，因为它们独特的多能性和无限的增殖潜力。为了将这些细胞的可能性转化为科学和医学的进步，人们必须了解它们如何整合环境刺激，包括可溶性化学因子，细胞外基质蛋白，细胞间相互作用和生物物理力量，在自我更新和分化决定中。该项目将具体解决如何在空间和时间上对微环境中的化学定义的文化系统的背景下的机械线索的ESC。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ESC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将被测试。 所采用的方法涉及三项综合活动的反复循环。首先，主要研究者（PI）在材料和界面方面的专业知识将用于设计和表征新型培养系统，该系统允许将空间和时间定义的机械应力应用于ESC集落。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用于构建数学模型，描述调控ESC分化的信号和途径的复杂性。这些模型将预测微环境线索对细胞的最佳呈现，以促进自我更新或分化;然后，这些预测将通过实验进行测试。来自这些实验的数据将提供额外的信息，这将促进进一步的模型完善。从该项目中获得的环境信号调节ESC研究之间的关系的科学理解，可能会提供一个合理的基础设计的ESC培养系统，这将有助于他们在研究和临床环境中的使用。该项目还将支持一项新的公众宣传计划，包括讨论干细胞生物学和工程技术方面的论坛以及ESC研究的社会影响。这一项目还将推动教育举措，招收高中生和本科生，特别是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。最后，该项目将支持PI制作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.