The Caco-2 human colon cancer cell line was obtained from the American Type Culture Collection (ATCC HTB-37TM, Manassas, VA, USA). These cells were routinely grown in 75 cm2 culture flasks (Corning, Amsterdam, NL) in high-glucose (25 mM) Dulbecco's Modified Eagle's medium (DMEM-glucose, Gibco, Breda, NL) supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin (Gibco), 1 mM Na-pyruvate (Gibco), 2 mM glutamax (Gibco), 25 mM HEPES buffer (Gibco) and 10% heat-inactivated (56°C, 45 min) fetal bovine serum (FBS, HyClone, Thermo Scientific, Breda, NL). Caco-2 cells were cultured at 37°C in a humidified atmosphere with 5% CO2 up to 80% confluency and then either subcultured or used for the experiments. For all experiments, Caco-2 cells with a passage number between 30 and 36 were proliferated or differentiated using either complete supplemented DMEM-glucose or complete supplemented DMEM-galactose [(= glucose-free DMEM (GIBCO) supplemented with 25 mM galactose as well as the other supplements as in DMEM-glucose)].
JanssenDuijghuijsen L.M., Grefte S., de Boer V.C., Zeper L., van Dartel D.A., van der Stelt I., Bekkenkamp-Grovenstein M., van Norren K., Wichers H.J, & Keijer J. (2017). Mitochondrial ATP Depletion Disrupts Caco-2 Monolayer Integrity and Internalizes Claudin 7. Frontiers in Physiology, 8, 794.
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