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03222022-Electrical Impedance for non-destructive, real time measurement of neural cell viability on microelectrode Arrays (Abstract for SOT 22)
Citation:
Choo, S., K. Wallace, T. Freudenrich, AND T. Shafer. 03222022-Electrical Impedance for non-destructive, real time measurement of neural cell viability on microelectrode Arrays (Abstract for SOT 22). Society of Toxicology 61st Annual Meeting and ToxExpo 2022, San DIego, California, March 27 - 31, 2022. https://doi.org/10.23645/epacomptox.19415969
Impact/Purpose:
Presentation to the Society of Toxicology 61st Annual Meeting and ToxExpo March 2022. Development of New Approach Methodologies are important for neurotoxicity and developmental neurotoxicity. This abstract summarizes an initial effort to improve the assessment of cytotoxicity in NAMs for neurotoxicity and DNT that are based on microelectrode assays.
Description:
Multielectrode arrays (MEA) are used to characterize chemical effects on neurotoxicity in vitro as they capture compound effects on neural network activity. Following assessment of compound effects on network function, cell viability is typically determined with cytotoxicity assays such as CellTiter Blue (CTB) and lactate dehydrogenase (LDH). Thus, tracking network activity and cell viability concomitantly is not possible. Recently, it has become possible to measure electrical impedance of cells attached to the electrodes, and this may serve as a surrogate measure for cell viability. In the present study, we sought to determine if impedance measurements can provide a contemporaneous measure of cell viability on MEAs. Thus, we compared viability assessment measured by impedance with that of LDH and CTB assays. Primary cultures from rat cortex plated on CytoView MEA 48 well plates were exposed to tween 20, tributyltin, deltamethrin, glyphosate, lindane and rotenone (0.03 - 30µM) on DIV 12 or 13. Impedance measurements were taken before treatment and at 0, 0.5, 1, 2, 4, 6, 24, 48 and 72h post treatment. After the last impedance measurement, CTB and LDH assays were performed for comparison. The acute compound effects on spontaneous network activity (number of active electrodes, nAE and weighted mean firing rate, wMFR) were also collected 0.5h post treatment for 0.5h. Glyphosate and lindane elicited no effect on the impedance measurement throughout the 72h of treatment. Tween 20, tributyltin, deltamethrin and rotenone caused biphasic changes in impedance in a time and concentration-dependent manner. Changes at 30µM tween 20 increased up to 2h and decreased thereafter, while 30µM deltamethrin increased impedance up to 24h and decreased thereafter. All concentrations of tributyltin tested increased impedance up to 6h and decreased after that, while rotenone increased impedance up to 24h and decreased thereafter. The CTB and LDH data conform to the impedance measurement to a greater extent, with CTB showing more noise. Tween 20, tributyltin and rotenone decreased both nAE and wMFR to 0% at highest concentration while glyphosate caused no or little effect on wMFR. Deltamethrin increased wMFR up to 0.3µM and decreased both nAE and wMFR thereafter while lindane increased wMFR up to 10µM and decreased nAE and wMFR thereafter. These results demonstrated impedance as a reliable cell viability measure. This abstract does not reflect EPA policy
URLs/Downloads:
DOI: 03222022-Electrical Impedance for non-destructive, real time measurement of neural cell viability on microelectrode Arrays (Abstract for SOT 22)
SOT 2022_CHOO_FINALFINAL.PDF (PDF, NA pp, 721.721 KB, about PDF)