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.
Copyright 2000 Academic Press.