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

基本信息

批准号：
9206500
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
项目类别：
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/9206500
关键词：
Acoustics Actins Affect Apoptosis Biological Assay Biomechanics Biophysics Cell Adhesion Cell Count Cell Death Cell Maintenance Cell Survival Cell physiology Cells Clone Cells Cloning Cytometry Cytoskeleton Degenerative Disorder Developmental Process Diabetes Mellitus Differentiation and Growth Disease model Dissociation E-Cadherin Future Gene Transfer Generations Goals Growth Investigation Laboratory culture Liquid substance Magnetism Mechanical Stimulation Mechanics Methods Microbubbles Microspheres Molecular Myosin Type II Pathologic Processes Physiologic pulse Physiological Processes Play Preclinical Drug Evaluation Process Protocols documentation Receptor Cell Regenerative Medicine Regulation Research Research Project Grants Resolution Resources Role Signal Transduction Source Spinal cord injury Stem cells Stretching Survival Rate Techniques Technology Therapeutic Tissues Translations Ultrasonography adhesion receptor base cell behavior cell growth experimental study extracellular genetic manipulation high throughput screening human embryonic stem cell human pluripotent stem cell improved induced pluripotent stem cell innovation insight laser tweezer mechanical force mechanotransduction migration new technology novel pluripotency polymerization practical application protein expression public health relevance regenerative therapy response spatiotemporal technology development tool

项目摘要

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（由申请人提供）：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应用的工具。这项研究的目标是开发一种基于超声波的新型技术，即声镊细胞术（ATC），作为强大的细胞力学和机械生物学。我们将进行系统和全面的研究，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和科学研究产生广泛的影响。证明 ATC 作为一种新颖实用的干细胞应用策略的实用性，特别是在人类多能干细胞 (hPSC) 维护和理解 hPSC 的机械生物学方面取得新进展，能够自我复制，同时保留产生任何细胞的能力。 hPSC 是一种特殊的细胞，为疾病建模、药物筛选和未来基于细胞的疗法治疗糖尿病和脊髓损伤等退行性疾病提供了有前景的来源，但控制 hPSC 的生长仍然具有挑战性，因为。目前克隆生长 hPSC 的方法效率低下，且对于基因操作和治疗目的的定义不明确，hPSC 在细胞分离和解离时容易发生细胞凋亡，解离的单个 hPSC 的克隆效率通常 < 1% 因此，我们提出以下具体目标。在这项研究中：1) 开发一个创新的 ATC 技术平台，用于应用时空控制的亚细胞机械力；2) 确定 ATC 对 hPSC 存活和克隆效率的影响； 3）揭示ATC刺激提高hPSC存活率和克隆效率的机制。