Robust microdroplet-based mechanical probes for wide-ranging mechanobiology applications

坚固的基于微滴的机械探针，适用于广泛的机械生物学应用

• 批准号: 10242779
• 负责人: Otger Campas
• 金额: $ 43.48万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242779
• 关键词: 3-Dimensional; Adult; Affect; Architecture; Benchmarking; Biochemical; Biological; Biological Models; Cell Communication; Cell Culture Techniques; Cell Surface Receptors; Cells; Cellular Spheroids; Chemicals; Chemistry; Communities; Cues; Custom; Defect; Development; Disease; Disease Progression; Embryo; Embryonic Development; Ensure; Environment; Fibrosis; Fluorocarbons; Goals; Grant; Homeostasis; Image; Impairment; In Situ; In Vitro; Knowledge; Label; Magnetic nanoparticles; Magnetism; Maintenance; Malignant Neoplasms; Measurement; Measures; Mechanical Stress; Mechanics; Methodology; Methods; Oils; Organ; Organoids; Outcome; Oxides; Performance; Physiological; Play; Positioning Attribute; Process; Property; Receptor Cell; Reporting; Reproducibility; Role; Signal Transduction; Solubility; Stress; Surface; System; Techniques; Technology; Testing; Time; Tissues; Transducers; Visualization; Work; base; behavior in vitro; biological systems; cell behavior; commercialization; cyanine dye; ferrite; ferrofluid; fluorophore; in vivo; innovation; interfacial; magnetic field; malignant phenotype; mechanical force; nanoparticle; new technology; novel; organ growth; prevent; sensor; stem cell differentiation; surfactant; technology development; three dimensional cell culture; tool; tumor; tumor progression

PROJECT SUMMARY Mechanical cues critically affect cell behaviors that are central to embryonic development, organ formation and the maintenance of tissue architecture and homeostasis. Both mechanical forces and the material properties of the cellular microenvironment (e.g., stiffness) are known to direct stem cell differentiation as well as alter the progression of malignant phenotypes during tumor progression. While it is generally acknowledged that mechanical forces and the material properties of the cellular microenvironment play a key role in the control of cell behaviors in vitro, the lack of technologies to perform quantitative measurements of mechanics in 3D cellular microenvironments has considerably hindered our ability to understand the role of mechanics in more physiologically relevant environments. PI Campas and coworkers have recently reported a novel methodology that enables direct in vivo and in situ mechanical measurements within 3D cellular microenvironments, including living tissues, for the first time. While groundbreaking, these methods, which make use of fluorescently-labeled, magnetically-responsive microdroplets of perfluorinated oil as mechanical sensors and actuators, remain strongly limited in scope because of current chemistry used to prepare the microdroplets. The current finicky chemical composition of the microdroplets strongly limits the scope of the technique, hampers the reproducibility of the measurements and precludes the dissemination of these methods to the wide biological and biomedical communities. In this multi-PI technology development grant, Campas, Sletten, and Zink team up to solve the existing problems with the microdroplet technology by developing new robust chemistries, including fluorinated surfactants, fluorophores, and magnetic nanoparticles, to enable accurate mechanical measurements with microdroplets in a wide range of biological systems, from living tissues and organs to organoids, tumors and 3D cell culture. In Aim 1, we will develop surfactants and fluorophores to properly control the droplet’s interfacial tension, the cell droplet interactions and their visualization in 3D multicellular systems. In Aim 2, we will develop robust perfluorocarbon-based ferrofluids with controlled interfacial tension, cell-droplet interactions and with strong magnetic properties enabling the application of larger forces. In Aim 3, we will test the functionality of the newly synthesized compounds and assess their performance in mechanical measurements in well-established 3D multicellular systems, both in vitro and in vivo. Upon completion of these aims, we will achieve an optimized microdroplet technology that is ready for commercialization and can be used in a wide range of systems, including living tissues, organoids, embryoid bodies, tumors and 3D cell culture, thereby making accessible these new tools for the study of mechanical cues (mechanobiology) in vivo to the entire biological and biomedical communities and potentially transforming our understanding of biological systems.

项目概要 机械信号严重影响对胚胎发育、器官形成和发育至关重要的细胞行为。 组织结构和体内平衡的维持。机械力和材料特性 已知细胞微环境（例如硬度）可指导干细胞分化并改变 肿瘤进展过程中恶性表型的进展。虽然人们普遍认为 机械力和细胞微环境的材料特性在控制中起着关键作用 体外细胞行为，缺乏对 3D 力学进行定量测量的技术 细胞微环境极大地阻碍了我们理解力学在更多领域中的作用的能力 生理相关环境。 PI Campas 和同事最近报告了一种新颖的方法 能够在 3D 细胞微环境中直接进行体内和原位机械测量， 首次包括活体组织。这些方法虽然具有开创性，但利用了 荧光标记的、磁响应的全氟化油微滴作为机械传感器和 由于目前用于制备微滴的化学方法，致动器的范围仍然受到严格限制。 目前微滴的化学成分要求严格，严重限制了该技术的范围， 妨碍了测量的可重复性并妨碍了这些方法的传播 广泛的生物和生物医学界。在这项多 PI 技术开发资助中，Campas、Sletten、 和 Zink 合作，通过开发新的稳健的技术来解决微滴技术现有的问题 化学物质，包括氟化表面活性剂、荧光团和磁性纳米粒子，以实现准确的 在广泛的生物系统中使用微滴进行机械测量，这些微滴来自活组织和 器官到类器官、肿瘤和 3D 细胞培养。在目标 1 中，我们将开发表面活性剂和荧光团 正确控制液滴的界面张力、细胞液滴相互作用及其 3D 可视化 多细胞系统。在目标 2 中，我们将开发鲁棒的全氟化碳基铁磁流体，并具有可控的 界面张力、细胞-液滴相互作用以及强磁性使应用成为可能 更大的力量。在目标 3 中，我们将测试新合成化合物的功能并评估其 在成熟的 3D 多细胞系统中，体外和体内的机械测量性能 体内。完成这些目标后，我们将实现优化的微滴技术，为 商业化，可用于广泛的系统，包括活组织、类器官、胚状体 身体、肿瘤和 3D 细胞培养，从而使这些新工具能够用于机械研究 体内对整个生物和生物医学界的线索（机械生物学），并有可能改变 我们对生物系统的理解。