Measuring Epithelial Barrier Integrity

Transepithelial resistance measures the integrity of cellular tight junctions and is a suitable method to be used in cell culture monolayers for the purpose of measuring the effect of bacteria during infection in vitro (Srinivasan et al., 2015 (link)). This methodology measures the electrical resistance, in ohms, as a measure of cellular barrier integrity. For the purpose of TEER measurement, the HCT-8 cells were grown on 0.4 μm and 12 mm pore size transwell inserts (Corning) and selected based on the formation of a confluent monolayer. Our aim was to investigate the effect of Auranta 3001 on the barrier properties of HCT-8 cells by taking TEER measurements at 3 h postinfection (±0.1 and 0.5% Auranta 3001) using an EVOM X meter connected to an Endohm chamber (World Precision Instruments).

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Sima F., Stratakos A.C., Ward P., Linton M., Kelly C., Pinkerton L., Stef L., Gundogdu O., Lazar V, & Corcionivoschi N. (2018). A Novel Natural Antimicrobial Can Reduce the in vitro and in vivo Pathogenicity of T6SS Positive Campylobacter jejuni and Campylobacter coli Chicken Isolates. Frontiers in Microbiology, 9, 2139.

Publication 2018

pore size transwell inserts EVOM X meter Endohm chamber

Bacteria Cell culture Cells Cellular junctions Electrical resistance Infection

Corresponding Organization : Univesity of Life Science "King Mihai I" from Timisoara

Other organizations : University of Bucharest, London School of Hygiene & Tropical Medicine

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Variable analysis

independent variables

Auranta 3001 concentration (0.1% and 0.5%)

dependent variables

Transepithelial electrical resistance (TEER) of HCT-8 cell monolayers

control variables

HCT-8 cell monolayers grown on 0.4 μm and 12 mm pore size transwell inserts
Time of TEER measurement (3 h postinfection)
EVOM X meter connected to an Endohm chamber (World Precision Instruments) used for TEER measurements

controls

No explicit mention of positive or negative controls

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