Trans‐endothelial electrical resistance was measured by using epithelial voltmeter (World Precision Instruments, FL, USA). In brief, after respective treatment, media were aspirated from the upper chamber and cell culture inserts were transferred to a new well and washed with phenol‐red free DMEM. TEER was analyzed with an epithelial voltmeter (World Precision Instruments, FL, USA), and the values were expressed as Ω*cm2. According to previous findings (6, 9, 11), when the TEER value reached over 300 Ω*cm2, the BBB model was considered validly established.
Epithelial voltmeter
Manufactured by World Precision Instruments
Sourced in United States
The Epithelial Voltmeter is a laboratory instrument designed to measure the electrical potential across epithelial cell layers. It provides precise and reliable measurements of the transepithelial voltage, which is a key parameter in the study of epithelial cell function and transport mechanisms.
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4 protocols using epithelial voltmeter
1
Measuring Endothelial Barrier Integrity
2
Measuring RPE Cell Barrier Function
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For the measurement of TEER, the RPE cells were grown on matrigel-coated transwell membranes. A membrane without cells was used as a background control. After 4–5 weeks of culture, TEER of the RPE cells was measured using an epithelial voltmeter (World Precision Instruments) and calculated by subtraction of background TEER from the control membrane. The TEER per unit area (Ω.cm2) was obtained by multiplying the raw TEER value by the surface area of the transwell membrane (0.33 cm2).
3
Evaluating Epithelial Barrier Integrity
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The epithelial barrier function in vitro was examined by trans-epithelial electrical resistance (TEER) using a 24-well transwell plate. The Caco-2 cells were seeded at a density of 2 × 104 cells/well in 24-well transwell insert (pore size 0.4 μm, Corning). After 48 h of culture, the TEER was measured with an epithelial voltmeter (World Precision Instruments, United States), and then the cells were treated with GA (10 μM). 2 h later, cells were treated with LPS (2 μg/mL). The TEER was measured after LPS and GA co-treatment for 2, 4, 6, and 24 h, respectively. The resistance measurements of cell-free filters (blank resistance) were subtracted from that seeded with cells. The resistance value of the filters and fluids was be subtracted in the calculation for each measurement.
4
TEER Measurement of Caco-2 Monolayer Irradiation
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The trans-epithelial electrical resistance (TEER) of Caco-2 monolayer formation was assessed using EVOM2 (link) Epithelial Voltmeter (World Precision Instruments) after 19–21 days in culture. It has established in the literature that Caco-2 cells can be used for permeabilization TEER measurements only when TEER values range between 300–600 Ω/cm2 (link).
Cells were treated with whey protein 150 μg/ml whey protein, in addition to different concentrations of GOP (50, 150 and 200 μg/ml) for 24 hours. After the initial assessment of TEER values, cells were subsequently irradiated with 2 Gy X-ray, with a dose rate of 1.14 Gy/min at room temperature, using FAQ RS-2000 Irradiator (Rad Source Technologies) in Dana-Farber Cancer Institute (Boston, USA). The environmental/procedural conditions of the sham-irradiated cells were similar to the irradiated cells. TEER measurements was performed initially before the irradiation and hourly for the first 6 hours post-irradiation, then every 3 hours till 48 hours, in presence/absence of different concentrations of GOP or whey protein.
Cells were treated with whey protein 150 μg/ml whey protein, in addition to different concentrations of GOP (50, 150 and 200 μg/ml) for 24 hours. After the initial assessment of TEER values, cells were subsequently irradiated with 2 Gy X-ray, with a dose rate of 1.14 Gy/min at room temperature, using FAQ RS-2000 Irradiator (Rad Source Technologies) in Dana-Farber Cancer Institute (Boston, USA). The environmental/procedural conditions of the sham-irradiated cells were similar to the irradiated cells. TEER measurements was performed initially before the irradiation and hourly for the first 6 hours post-irradiation, then every 3 hours till 48 hours, in presence/absence of different concentrations of GOP or whey protein.
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