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

背景：在这个项目中，？干细胞？术语“成体干细胞”仅指非胚胎来源的成体干细胞（也称为体干细胞）。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成特定的后代。它们将从国家非营利生物资源中心（例如ATCC）通常可获得的现有细胞系中获得。成人干细胞的来源是什么？浪费？手术切除后丢弃的组织，由转化研究中心和医疗机构提供。技术描述：该项目将提供干细胞如何耐受流体流动的剪切应力的基础工程和生物学理解。它将研究不同制度下的剪切应力，从血管和间质流的生理条件下的干细胞聚集体在体外条件下的移液管研磨和混合叶轮。将使用比较和定量蛋白质组学分析评价剪切应力对干细胞球聚集体的影响，这将允许阐明潜在的机械转导分子特征和信号传导途径。此外，还将对天然抗剪切血液细胞进行蛋白质组学分析，以确定关键的抗剪切基因，并将这些基因工程化到模型干细胞中，以提高它们在高剪切条件下的存活率。拟议的工作解决了一个重要但相对研究不足的研究领域，并将通过创造性地结合使用工程和分子生物学工具来显着推进该领域。更广泛的意义和重要性：拟议的项目本质上是跨学科的，将为生物加工、生物工程和机械生物学等广泛领域做出根本性的概念验证贡献。特别是，该项目将研究如何在剪切应力条件下保持干细胞的高活力。为细胞治疗应用产生可观数量的干细胞将不可避免地需要使用大型容器，其中用叶轮混合产生高剪切环境。为了提高干细胞在这种条件下的生存能力，本项目将研究模型剪切应力环境中的干细胞，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之外，这项工作的发现还将对理解其他细胞（如循环肿瘤细胞）如何在剪切应力条件下存活产生重要影响，并允许开发新的靶向药物。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：为了扩大非洲裔美国学生在工程研究中的参与，PI将与区域HBCU和亚拉巴马？黑带？靠近亚拉巴马大学的高中。这种互动将包括为该地区的这些学生定期举办研讨会和提供实践经验的机会。为了扩大妇女在工程研究中的参与，PI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研究所本科夏季研究计划和亚拉巴马大学招募这两个代表性不足群体的学生研究人员。工程学夏令营学生介绍。这项研究已通过工程招标，这是工程教育和中心司的工程计划的扩大参与的一部分，扩大参与研究启动赠款资助。

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.