Sensitivity and validation of porous membrane electrical cell substrate impedance spectroscopy (PM-ECIS) for measuring endothelial barrier properties

• DOI: 10.1101/2023.07.11.548631
• Publication Date: 2023-07-13T03:56:29Z
• Authors: Alisa Ugodnikov; Oleg Chebotarev; Henrik Persson; Craig A. Simmons
• Keywords: 0303 health sciences; 03 medical and health sciences; Dielectric Spectroscopy; Electric Impedance; Human Umbilical Vein Endothelial Cells; Humans; Endothelial Cells; Membranes, Artificial; Porosity; Electrodes

ABSTRACT Conventional trans-endothelial electrical resistance (TEER) setups are invasive and cannot directly measure monolayer integrity in co-culture. These limitations addressed by porous membrane cell-substrate impedance sensing (PM-ECIS), which measures barrier cell monolayers grown on permeable membranes patterned with electrodes. Here we advance the design utility of PM-ECIS investigating its sensitivity to working electrode size correlation TEER. Gold electrodes were fabricated inserts using hot embossing UV lithography, diameters 250, 500, 750 µm within same insert. Frequency scans confluent primary endothelial showed normalized peaking at 4 kHz, consistent traditional solid substrate ECIS. Sensitivity changes (4 kHz) during formation was inversely proportional size, smallest being most sensitive (p<0.001). Similarly, smaller (40 corresponding spreading proliferation Barrier disruption EGTA detectable 250 (p<0.01), 500 (p=0.058). Resistances measured vs. TEER for sodium chloride solutions positively significantly correlated all sizes (r>0.9; p<0.0001), but only (r=0.71; p=0.058). data inform selection specific applications, support as a promising alternative conventional due capacity direct, non-invasive, real-time assessment cells cultured membranes.