Acoustic tweezing cytometry: technology development and stem cell applications

声学镊子细胞术：技术开发和干细胞应用

• 批准号: 9049494
• 负责人: CHERI X DENG
• 金额: $ 48.45万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-15 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9049494
• 关键词: Acoustics; Actins; Affect; Apoptosis; Biological Assay; Biomechanics; Biophysics; Cell Adhesion; Cell Count; Cell Death; Cell Maintenance; Cell Survival; Cell physiology; Cells; Cloning; Cytometry; Cytoskeleton; Degenerative Disorder; Developmental Process; Diabetes Mellitus; Differentiation and Growth; Disease model; Dissociation; E-Cadherin; Future; Gene Transfer; Generations; Goals; Growth; Health; Human; Investigation; Laboratory culture; Life; Liquid substance; Magnetism; Mechanical Stimulation; Mechanics; Methods; Microbubbles; Microspheres; Molecular; Myosin Type II; Pathologic Processes; Physiologic pulse; Physiological Processes; Play; Pluripotent Stem Cells; Preclinical Drug Evaluation; Process; Protocols documentation; Receptor Cell; Regenerative Medicine; Regulation; Research; Research Project Grants; Resolution; Resources; Role; Signal Transduction; Source; Spinal Cord; Spinal cord injury; Stem cells; Stretching; Survival Rate; Techniques; Technology; Therapeutic; Tissues; Translations; Ultrasonography; adhesion receptor; base; cell behavior; cell growth; extracellular; genetic manipulation; high throughput screening; human embryonic stem cell; improved; induced pluripotent stem cell; innovation; insight; laser tweezer; meetings; migration; new technology; novel; pluripotency; polymerization; practical application; protein expression; regenerative therapy; research study; response; technology development; tool

 DESCRIPTION (provided by applicant): Mechanosensitivity to extracellular mechanical signals is central to many developmental, physiological, and pathological processes, affecting cell functions including growth, migration, differentiation, and apoptosis. Understanding the molecular mechanisms underlying mechanotransduction process rely on tools capable of applying controlled mechanical forces to cells to elicit and assess cellular responses. The goal of this research is to develop a novel ultrasound-based technology, acoustic tweezing cytometry (ATC), as a powerful cell mechanics and mechanobiology tool. We will perform systematic and comprehensive studies to develop innovative ATC platform and characterize subcellular force generation in ATC for mechanical regulation of cells, which will have broad impact on many practical applications as well as scientific investigations. In this research, we will develop and demonstrate the utility of ATC as a novel and practical strategy for stem cell applications, specifically to enable novel advances in human pluripotent stem cell (hPSC) maintenance and understanding of mechanobiology of hPSCs. Capable of replicating themselves while retaining the ability to give rise to any type of specialized cells, hPSCs provide promising sources for disease modeling, drug screenings, and future cell-based therapeutics to treat degenerative diseases such as diabetes mellitus and spinal cord injury. However, controlling hPSC growth remains challenging because present methods to clonally grow hPSCs are inefficient and poorly defined for genetic manipulation and therapeutic purposes. hPSCs are vulnerable to apoptosis upon cellular detachment and dissociation, with a cloning efficiency of dissociated single hPSCs generally < 1%. Therefore, we propose the following specific aims in this research: 1) to develop an innovative ATC technology platform for applying spatiotemporally controlled subcellular mechanical forces; 2) to determine the effects of ATC on the survival and cloning efficiency of hPSCs; and 3) to reveal the mechanisms of ATC stimulation for improving survival and cloning efficiency of hPSCs.

 描述(申请人提供)：对细胞外机械信号的机械敏感性是许多发育、生理和病理过程的核心，影响细胞的功能，包括生长、迁移、分化和凋亡。了解机械转导过程背后的分子机制依赖于能够对细胞施加受控机械力的工具来诱导和评估细胞反应。本研究的目标是开发一种新的基于超声的技术--声镊子细胞术(ATC)，作为一种强大的细胞力学和机械生物学工具。我们将开展系统和全面的研究，以开发创新的ATC平台，并表征ATC中用于细胞机械调节的亚细胞力产生，这将对许多实际应用以及科学研究产生广泛影响。在这项研究中，我们将开发和 证明ATC作为干细胞应用的一种新颖和实用的策略的实用性，特别是使人类多能干细胞(HPSC)维护和理解hPSC的机械生物学方面的新进展成为可能。HPSC能够在自我复制的同时保持产生任何类型的特化细胞的能力，为疾病建模、药物筛选以及未来基于细胞的治疗退行性疾病(如糖尿病和脊髓损伤)提供了有前景的来源。然而，控制hPSC的生长仍然具有挑战性，因为现有的克隆培养hPSC的方法效率低下，并且对于遗传操作和治疗目的的定义不明确。HPSCs在细胞分离和分离后容易发生凋亡，其克隆率一般为1%。因此，我们在本研究中提出了以下具体目标：1)开发一种创新的应用时空可控亚细胞机械力的ATC技术平台；2)研究ATC对hPSCs存活和克隆效率的影响；3)揭示ATC刺激提高hPSCs存活和克隆效率的机制。