Measurement of EGTA-Mediated Barrier Impairment by Impedance Spectroscopy

Abstract

For the widespread use of in vitro models of barrier tissues in practice, it is necessary to develop approaches to the automation of cultivation and monitoring of the state of cells. Impedance spectroscopy is a promising and informative method for assessing the state of cell barriers. In this work, the dynamics of electrical parameters in an in vitro model of the intestinal barrier with a high temporal resolution has been studied using impedance spectroscopy. For the first time, the detailed kinetics of changes in TEER, electrical capacitance and additional resistance of a monolayer of Caco-2 cells at various EGTA concentrations was recorded. It was demonstrated that spontaneous restoration of the integrity of the cell barrier is possible in the presence of 2 mM EGTA in the medium, which begins approximately 6 h after the compound addition. A strong increase in TEER was also found immediately after the addition of EGTA, which probably depended on the EGTA concentration and can be explained by a decrease in intracellular conductance due to the closure of transmembrane channels. impedance spectroscopy, intestine, TEER, electrical capacitance, EGTA The study was funded by the Russian Science Foundation (project 16-19-10597).