Study of the Electrical Characteristics of Epithelial Cell Layers for the Development of an Organ-on-a-Chip System
Gorbachev I.A., IRE Kotelnikova RAS
Zhukova O.A.,Federal State Budgetary Institution “Research Institute of Pulmonology” of the Federal Medical-Biological Agency of Russia
Baklaushev V.P., Federal State Budgetary Institution “Research Institute of Pulmonology” of the Federal Medical-Biological Agency of Russia
Zikov K.A., Federal State Budgetary Institution “Research Institute of Pulmonology” of the Federal Medical-Biological Agency of Russia
Kuznetsova I.E., IRE Kotelnikova RAS
Kolesov V.V., IRE Kotelnikova RAS
Abstract
Transepithelial electrical resistance is a key parameter used to assess the functional state of a cell layer. Its monitoring is particularly important in the development of organ-on-a-chip systems. The aim of this study was to evaluate the electrical characteristics of a Caco-2 cell monolayer formed on a permeable membrane in a two-chamber microfluidic cell.
The microfluidic device was fabricated using 3D photopolymer printing. The device consisted of two halves containing a system of microchannels separated by a permeable membrane with a working area 5 mm in diameter and a chamber height of 3 mm. An electrospun membrane made of spidroin-modified polycaprolactone was used as the permeable partition. A Caco-2 cell layer was formed on the membrane surface. Indirect electrical contact with the cell layer through the conductive liquid medium was provided by silver electrodes. Cell culture medium was used as the electrolyte. Impedance measurements were performed using a P-45X potentiostat–galvanostat over a frequency range from 1 Hz to 10 kHz with an AC perturbation amplitude of 0.01 V. The electrical characteristics of the cell layer were determined during the study.
The experiments were performed using a cell containing a cell-free membrane and after the formation of a cell layer on the membrane surface. The presence of the cell monolayer shifted the Nyquist plot toward higher resistance values. To interpret the obtained results, an equivalent electrical circuit was proposed. The circuit included the resistance of the liquid medium, constant phase elements, and parallel resistor–capacitor elements describing the electrical properties of the membrane and the cell layer. Distribution of relaxation times analysis was applied to identify individual electrochemical processes, while the quantitative parameters of the equivalent-circuit elements were determined by fitting the experimental impedance spectra to the selected electrical model.
The study was supported by the Russian Science Foundation, Grant No. 25-19-00872.
Speaker
Gorbachev Ilya A.
IRE Kotelnikova RAS
Russia
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