Abstract
Electric cell–substrate impedance sensing (ECIS) has been used as a real-time impedance-based method to quantify cell behavior in tissue culture. The method is capable of measuring both the resistance and capacitance of a cell-covered microelectrode at various AC frequencies. In this study, we demonstrate the application of high-frequency capacitance measurement (f = 40 or 64 kHz) for the sensitive detection of both the micromotion and wound-healing migration of human mesenchymal stem cells (hMSCs). Impedance measurements of cell-covered electrodes upon the challenge of various concentrations of carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), from 0.1 to 30 μM, were conducted using ECIS. FCCP is an uncoupler of mitochondrial oxidative phosphorylation (OXPHOS), thereby reducing mitochondrial ATP production. By numerically analyzing the time-series capacitance data, a dose-dependent decrease in hMSC micromotion and wound-healing migration was observed, and the effect was significantly detected at levels as low as 0.1 μM. While most reported works with ECIS use the resistance/impedance time series, our results suggest the potential use of high-frequency capacitance time series for assessing migratory cell behavior such as micromotion and wound-healing migration.
Highlights
Many animal cell types exhibit the ability to move from one location to another and carry out other subtle motions
A wound or gap is created on the surface of a confluent monolayer of cells through mechanical, electrical, chemical, or thermal means, and the wound or gap closure is observed by optical microscopy under a variety of experimental conditions [6]
To monitor human mesenchymal stem cells (hMSCs) attachment and spreading, the frequency-dependent capacitances of the cell-covered electrodes were followed at various frequencies
Summary
Many animal cell types exhibit the ability to move from one location to another and carry out other subtle motions. To measure the two-dimensional cell migration of adherent cells, the most common and straightforward approach is using the wound-healing assay In this method, a wound or gap is created on the surface of a confluent monolayer of cells through mechanical, electrical, chemical, or thermal means, and the wound or gap closure is observed by optical microscopy under a variety of experimental conditions [6]. A wound or gap is created on the surface of a confluent monolayer of cells through mechanical, electrical, chemical, or thermal means, and the wound or gap closure is observed by optical microscopy under a variety of experimental conditions [6] These visual observations can be graded to provide an estimate of the rate of cell migration. The ECIS method has been successfully applied to investigate wound-healing migration of various cell cultures including epithelia, endothelia, keratinocyte, and mesenchymal stem cells [7,8,9,10]
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