BRIGE: Effects of Varying Fluid Shear Stress on Stem Cell Sphere Aggregates

背景：在这个项目中，干细胞？仅指非胚胎来源的成体干细胞(也称为体细胞)。与所有类型的干细胞一样，这些成体干细胞能够自我更新，并能够分化为特定的后代。它们将从国家非营利性生物资源中心(如ATCC)普遍提供的现有细胞系中获得。成体干细胞的其他来源将来自？废物？手术切除后被丢弃的组织，由转化研究中心和医疗机构提供。技术描述：该项目将为干细胞如何耐受流体流动产生的剪应力提供基本的工程学和生物学理解。它将研究不同剪应力条件下的干细胞聚集体，范围从血管和间质流动的生理条件到吸管摩擦和混合叶轮的体外条件。剪切应力对干细胞球团聚体的影响将通过比较和定量蛋白质组学分析来评估，这将有助于阐明潜在的机械转导分子特征和信号通路。此外，还将对天然抗剪切血细胞进行蛋白质组学分析，以确定关键的抗剪切基因，这些基因将在模型干细胞中进行工程，以提高它们在高切变条件下的生存能力。拟议的工作涉及一个重要但相对研究较少的研究领域，并将通过创造性地结合使用工程学和分子生物学工具来显着推动该领域的发展。此外，该项目将通过深入了解干细胞在高剪切环境中的正确处理和生长，为再生医学领域做出革命性的贡献。BROADER意义和重要性：拟议的项目本质上是跨学科的，将对包括生物加工、生物工程和机械生物学在内的广泛领域做出基本的概念验证贡献。特别是，该项目将研究如何在剪切力条件下保持干细胞的高活性。为细胞治疗应用产生相当数量的干细胞将不可避免地需要使用大型容器，在大型容器中与叶轮混合将产生高剪切环境。为了提高干细胞在这种条件下的生存能力，该项目将在模型剪应力环境中研究干细胞，并阐明与其生存相关的潜在生物学机制。除了干细胞生物技术，这项工作的发现也将对理解其他细胞，如循环中的肿瘤细胞，如何在剪应力条件下生存，并允许新的靶向药物开发具有重要的影响。拟议研究的组成部分将被纳入核心本科化学工程课程。扩大参与活动：为了扩大非裔美国学生对工程研究的参与，PI将与地区HBCU和阿拉巴马州？黑带？距离阿拉巴马大学很近的高中。这种互动将包括为该地区的这些学生举办定期研讨会和亲身体验机会。为了让妇女更多地参与工程研究，国际工程师学会将继续与女工程师协会的成员互动。此外，PI将通过霍华德·休斯医学院本科生暑期研究计划和阿拉巴马大学S学生工程导论夏令营招募这两个代表性不足群体的学生研究人员。这项研究是通过工程学征集中的扩大参与研究启动补助金资助的，这是工程教育和中心司扩大参与工程计划的一部分。

BACKGROUND:In this project, ?stem cells? refer exclusively to adult stem cells (also known as somatic stem cells) of non-embryonic origins. As common in all types of stem cells, these adult stem cells are capable of self-renewal and are able to differentiate into specified progenies. They will be acquired from existing cell lines commonly available at national nonprofit biological resource centers (e.g. ATCC). Additional source of adult stem cells will be from ?waste? tissues that are discarded after surgical resection, provided by translational research centers and medical institutions.TECHNICAL DESCRIPTION:This project will provide fundamental engineering and biological understanding of how stem cells tolerate shear stress from fluid flow. It will investigate stem cell aggregates under varying regimes of shear stresses, ranging from physiological conditions of vascular and interstitial flows to in vitro conditions of pipette trituration and mixing impellers. The effects of shear stresses on stem cell sphere aggregates will be evaluated using comparative and quantitative proteomic analysis, which would allow the elucidation of underlying mechanotransduction molecular signatures and signaling pathways. In addition, proteomic analysis of natively shear-resistant hematological cells will be performed to identify key shear-resistant genes, and these will be engineered in model stem cells to confer them improved survivability in high shear conditions. The proposed work addresses an important yet relatively understudied area of research and will significantly advance the field by its creative combined use of engineering and molecular biology tools. Furthermore, the project is poised to provide transformative contributions to the field of regenerative medicine by providing insights into proper handling and growth of stem cells in high shear environment.BROADER SIGNFICANCE AND IMPORTANCE:The proposed project is inherently interdisciplinary and will make fundamental proof-of-concept contributions to a broad number of fields, including bioprocessing, bioengineering, and mechanobiology. In particular, the project will investigate how to retain high viability of stem cells in shear stress conditions. Generating an appreciable number of stem cells for cell therapy applications would inevitably require the use of large vessels, in which mixing with impellers creates high shear environment. To improve the survivability of stem cells in such conditions, this project will investigate stem cells in model shear stress environments and elucidate the underlying biological mechanisms that correlate to their survival. Beyond stem cell biotechnology, the findings of this work will also have important ramifications in understanding how other cells, such as circulating tumor cells, survive in shear stress conditions, and allow for novel targeted drug development. Components of the proposed research will be integrated into core undergraduate chemical engineering courses. BROADENING PARTICIPATION ACTIVITIES:To broaden participation of African-American students in engineering research, the PI will interact with regional HBCUs and the Alabama ?Black Belt? high schools that are in close proximity to the University of Alabama. This interaction will involve regular seminars and hands-on experience opportunities for these students in the region. To broaden the participation of women in engineering research, the PI will continue to interact with the members of the Society of Women Engineers. In addition, the PI will recruit student researchers of both underrepresented groups through the Howard Hughes Medical Institute Undergraduate Summer Research Program and through the University of Alabama?s Student Introduction to Engineering Summer Camp. This research has been funded through the Broadening Participation Research Initiation Grants in Engineering solicitation, which is part of the Broadening Participation in Engineering Program of the Engineering Education and Centers Division.