High-Throughput Single Cell Mechanomics

高通量单细胞力学

• 批准号: 10462589
• 负责人: Pranav Soman
• 金额: $ 20.08万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462589
• 关键词: 3D Print; Address; Algorithms; Atomic Force Microscopy; Behavior; Biochemical; Biomedical Research; Biophysics; Calcium; Calcium Oscillations; Calcium Signaling; Cell model; Cell physiology; Cells; Chemical Stimulation; Chemicals; Coin; Consequentialism; Data; Development; Dimensions; Flow Cytometry; Fluorescence Microscopy; Frequencies; Future; Gelatin; Gene Expression; Gene Proteins; Generations; Heterogeneity; Human; Hybrids; Hydrogels; Individual; Joints; Lasers; Light; Literature; Magnetism; Measures; Mechanical Stimulation; Mechanics; Metabolism; Methods; Microelectrodes; Microfluidic Microchips; Microfluidics; Microspheres; Mission; Modeling; National Center for Complementary and Integrative Health; National Institute of General Medical Sciences; Optics; Outcome; Periodicity; Pharmaceutical Preparations; Phenotype; Physical Stimulation; Population; Printing; Publications; Publishing; Regenerative Medicine; Research; Research Personnel; Resolution; Signal Transduction; Soman; Stains; Stimulus; Stretching; Suspensions; Technology; Testing; Time; Torsion; United States National Institutes of Health; Work; base; biophysical properties; cell preparation; cell type; design; heuristics; high risk; innovation; innovative technologies; laser tweezer; light intensity; light microscopy; mechanical stimulus; mesenchymal stromal cell; population based; progenitor; prospective; response; scale up; stem cell biology; technological innovation; tumor

Summary. Phenotypic heterogeneity in cellular bulk populations can result in consequential differences in their response to physical as well as biochemical stimuli. To assess heterogeneity at single cell resolution, several methods have been developed, yet true predictability of cells’ future behavior cannot be reliably determined. To address this challenge, the proposed work will develop a new technological approach to solve the bulk cell heterogeneity problem coined as ‘single cell mechanomics’. This technology will record compression induced dynamic signaling response of single cells to predict and/or drive their future behavior. The technological innovation consists of a ‘smart’ microfluidic device with light actuated microtraps that can capture and compress single cells, and concurrently assess their signaling response, before releasing and capturing each individual cells for subsequent downstream monoclonal culture and analysis. To prove feasibility of this technology, human mesenchymal stromal cells (MSCs) will be used as a representative mechanoresponsive and highly heterogeneous cell type. Aim 1 will design and develop ‘smart’ microfluidic devices with light-actuated mictraps, while Aim 2 will establish a framework to predict and/or drive single cells’ phenotypic outcome based on calcium oscillation dynamics of mechanically compressed single cells. Multivariate predictive analyses will be used to identify relationships between compressive stimuli, calcium signaling, and phenotypic outcome. New relationships derived from this work will be used to identify and sort target cell populations based on their future phenotypes. At present, there is no demonstration of such a technology in the literature. This aligns with the high-risk requirements of this R21 solicitation of having significant future impact.

总结 细胞群体的表型异质性可能导致其发生相应的差异。 对物理和生化刺激的反应为了评估单细胞分辨率的异质性，需要进行一些研究。 方法已经开发出来，但细胞未来行为的真正可预测性仍无法可靠地确定。 为了应对这一挑战，拟议的工作将开发一种新的技术方法来解决散装电池的问题 异质性问题被称为“单细胞力学”，该技术将记录压缩引起的。 单细胞的动态信号响应来预测和/或驱动它们的未来行为。 创新包括“智能”微流体装置，带有光驱动微陷阱，可以捕获和压缩 单细胞，并在释放和捕获每个个体之前同时评估其信号反应 细胞用于后续下游单克隆培养和分析，以证明该技术的可行性。 间充质基质细胞（MSC）将被用作具有代表性的机械反应性和高度 目标 1 将设计和开发带有光驱动微阱的“智能”微流体装置， 而目标 2 将建立一个框架来预测和/或驱动基于钙的单细胞表型结果 机械压缩的单细胞的振荡动力学将用于。 确定压缩刺激、钙信号传导和表型结果之间的关系 新。 从这项工作中得出的关系将用于根据目标细胞群的未来来识别和分类 目前，文献中还没有这种技术的证明。 本次 R21 征集的高风险要求对未来产生重大影响。

项目成果

Printing Double-Network Tough Hydrogels Using Temperature-Controlled Projection Stereolithography (TOPS).

• DOI: 10.1021/acsami.3c04661
• 发表时间: 2023-06-28
• 期刊: ACS APPLIED MATERIALS & INTERFACES
• 影响因子: 9.5
• 作者: Kunwar, Puskal;Andrada, Bianca Louise;Poudel, Arun;Xiong, Zheng;Aryal, Ujjwal;Geffert, Zachary J. J.;Poudel, Sajag;Fougnier, Daniel;Gitsov, Ivan;Soman, Pranav
• 通讯作者: Soman, Pranav