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利用数字化声流技术进行细胞-基质黏附分析。

Profiling Cell-Matrix Adhesion Using Digitalized Acoustic Streaming.

机构信息

Department of Intelligent Systems Engineering , Indiana University , Bloomington , Indiana 47405 , United States.

Medical Sciences Program , Indiana University School of Medicine , Bloomington , Indiana 47405 , United States.

出版信息

Anal Chem. 2020 Jan 21;92(2):2283-2290. doi: 10.1021/acs.analchem.9b05065. Epub 2020 Jan 9.

DOI:10.1021/acs.analchem.9b05065
PMID:31880433
Abstract

Profiling the kinetics of cell-matrix adhesion is of great importance to understand many physiological and pathological processes such as morphogenesis, tissue homeostasis, wound healing, and tumorigenesis. Here, we developed a novel digital acoustofluidic device for parallel profiling cell-matrix adhesion at single-cell level. By introduction of localized and uniform acoustic streaming into an open chamber microfluidic device, the adherent cells within the open chamber can be detached by the streaming-induced Stokes drag force. By digital regulation of pulsed acoustic power from a low level to high levels, the hundreds of adherent cells can be ruptured from the fibronectin-coated substrate accordingly, and their adhesive forces (from several pN to several nN) and kinetics can be determined by the applied power and cell incubation time. As a proof-of-concept application for studying cancer metastasis, we applied this technique to measure the adhesion strength and kinetics of human breast cancer cells to extracellular matrix such as fibronectin and compared their metastatic potentials by measuring the rupture force of cancer cells representing malignant (MCF-7 cells and MDA-MB-231 cells) and nonmalignant (MCF-10A cells) states. Our acoustofluidic device is simple, easy to operate, and capable of measuring, in parallel, hundreds of individual cells' adhesion forces with a resolution at the pN level. Thus, we expect this device could be widely used for both fundamental cell biology research as well as development of cancer diagnostics and tissue engineering technologies.

摘要

研究细胞-基质黏附的动力学对于理解许多生理和病理过程非常重要,例如形态发生、组织稳态、伤口愈合和肿瘤发生。在这里,我们开发了一种新颖的数字声流控装置,用于在单细胞水平上平行分析细胞-基质黏附。通过将局部且均匀的声流引入开放式腔微流控装置,可以通过流致斯托克斯曳力将开放式腔中的黏附细胞分离。通过数字调节从低水平到高水平的脉冲声功率,可以相应地将数百个黏附细胞从纤维连接蛋白涂覆的基底上破裂,并且可以通过施加的功率和细胞孵育时间来确定它们的黏附力(从几个皮牛顿到几个毫牛顿)和动力学。作为研究癌症转移的概念验证应用,我们应用该技术来测量人乳腺癌细胞与细胞外基质(如纤维连接蛋白)的黏附强度和动力学,并通过测量代表恶性(MCF-7 细胞和 MDA-MB-231 细胞)和非恶性(MCF-10A 细胞)状态的癌细胞的破裂力来比较它们的转移潜力。我们的声流控装置简单、易于操作,并且能够以皮牛顿级的分辨率并行测量数百个单个细胞的黏附力。因此,我们期望该装置能够广泛用于基础细胞生物学研究以及癌症诊断和组织工程技术的开发。

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