The human colon adenocarcinoma Caco-2 cell line (passage number, 18) was obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA). The cells were cultured in DMEM containing 4.5 g/L glucose, 10% FBS, antibiotics (100 units/mL penicillin and 100 μg/mL streptomycin), and 1% NEAA at 37 °C in a humidified atmosphere of 5% CO2 in air.
In the present study, Caco-2 cells plated at a passage number (passage number, 23–35) similar to that reported by Watari et al. [47 (link)] were cultured in 6- or 12-well transwell chambers (Corning, NY, USA) at a density of 4 × 104 cells/cm2, and the medium was replaced every other day until 21 days. The mean TEER value was 1528 ± 117 Ω·cm2 (Figure S2 ), which exceeded the TEER cut-off value of 300 Ω·cm2, as measured by a Millicell-ERS volt-ohm meter (Millipore, Temecula, CA, USA). The differentiated Caco-2 cells were then used for TEER measurement, permeability tracer flux assay, Western blot analysis, and immunofluorescent staining.
In the present study, Caco-2 cells plated at a passage number (passage number, 23–35) similar to that reported by Watari et al. [47 (link)] were cultured in 6- or 12-well transwell chambers (Corning, NY, USA) at a density of 4 × 104 cells/cm2, and the medium was replaced every other day until 21 days. The mean TEER value was 1528 ± 117 Ω·cm2 (