Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces
Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces
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DOI：
10.1006/excr.2000.4919
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发表时间：
2000-08-25
影响因子：
3.7
通讯作者：
Giaever, I
中科院分区：
文献类型：
--
作者：
Wegener, J;Keese, CR;Giaever, I
本文描述了一种称为细胞-基质阻抗传感 (ECIS) 的实验技术的优化，用于定量实时监测哺乳动物细胞的附着和扩散。该方法基于测量沉积在培养皿上并用作生长基质的小金膜电极的交流阻抗的变化。基于实验数据和理论考虑，我们证明高频电容测量（f = 40 kHz）最适合跟踪由于细胞扩散而增加的电极表面覆盖范围。该方法具有出色的时间分辨率，可以在各种实验条件下深入分析细胞扩散动力学。使用 ECIS，我们研究了上皮 MDCK 细胞（II 株）在不同蛋白质涂层上的附着和铺展，并研究了二价阳离子对铺展动力学的影响。我们量化了模拟纤连蛋白和其他细胞外基质蛋白（RGDS）识别序列的可溶性肽的抑制作用。我们还应用 ECIS 技术通过这些肽监测汇合成纤维细胞层 (WI38/VA-13) 的脱离。 (C) 2000 年学术出版社。
This article describes the optimization of an experimental technique referred to as electric cell-substrate impedance sensing (ECIS) to monitor attachment and spreading of mammalian cells quantitatively and in real time. The method is based on measuring changes in AC impedance of small gold-film electrodes deposited on a culture dish and used as growth substrate. Based on experimental data and theoretical considerations we demonstrate that high-frequency capacitance measurements (f = 40 kHz) are most suited to follow the increasing surface coverage of the electrode due to cell spreading. The excellent time resolution of the method allowed an in-depth analysis of cell spreading kinetics under various experimental conditions. Using ECIS we studied the attachment and spreading of epithelial MDCK cells (strain II) on different protein coatings, and investigated the influence of divalent cations on spreading kinetics. We quantified the inhibitory effect of soluble peptides that mimic the recognition sequence of fibronectin and other extracellular matrix proteins (RGDS). We also applied the ECIS technique to monitor the detachment of confluent fibroblastic cell layers (WI38/VA-13) by means of these peptides. (C) 2000 Academic Press.