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.

概括。细胞大量种群中表型异质性可能导致其结果差异 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，有几个 已经开发了方法，但是无法确定细胞未来行为的真正可预测性。到 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单元 异质性问题被认为是“单细胞机械学”。该技术将记录引起的压缩 单细胞的动态信号响应预测和/或推动其未来行为。技术 创新由一个“智能”微流体设备和光激活的微饰面组成，可以捕获和压缩 单个单元格，并同时评估其信号响应，然后释放和捕获每个人 细胞进行随后的下游单克隆培养和分析。为了证明这项技术的可行性，人类 间充质基质细胞（MSC）将用作代表性的机械响应，高度 异质细胞类型。 AIM 1将设计和开发带有光线片段的“智能”微流体设备， 而AIM 2将建立一个基于钙的框架来预测和/或驱动单细胞的表型结果 机械压缩单细胞的振荡动力学。多元预测分析将用于 确定压缩刺激，钙信号传导和表型结果之间的关系。新的 从这项工作中得出的关系将根据其未来来识别和分类目标细胞种群 表型。目前，文献中没有这样的技术。这与 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