Culture and Maintenance of Esophageal Cell Lines

Human esophageal epithelial cells, STR, were cultured as previously described. In brief, Keratinocyte-SFM medium was supplemented with 1 ng/mL Epidermal Growth Factor, 0.05 mg/mL Bovine Pituitary Extract, and 1% penicillin streptomycin antibiotics (Gibco™, for Life Technologies, Inc., Carlsbad, CA, USA). Bile/acid-tolerant STR cells were established by repeated exposure to a 100 μM of cocktail of bile salts (20 μM of each, deoxycholic acid (MP Biomedicals, Santa Ana, CA, USA), glycocholic acid, taurocholic acid, sodium glycodeoxycholate and sodium glycochenodeoxycholate (all Sigma, St. Louis, MO, USA) as reported) [34 (link)].The non-neoplastic Barrett’s epithelial cell lines CPA, CPB, BAR-T and BAR-10T cells were isolated from non-dysplastic metaplasia (kind gift from Dr. Rhonda Souza, Baylor Scott White) [35 (link)], and were cultured in Epithelial Cell Medium-2 supplemented with 5% epithelial cell growth supplement-2 (EpiCGS-2) and 5% penicillin/streptomycin antibiotics (ScienceCell™ Research Laboratories, Carlsbad, CA, USA). The esophageal adenocarcinoma cell lines, FLO1, OE33 and OE19 (kind gift of Dr. El-Rifai, University of Miami), were grown in RPMI with 10% FBS and 1% penicillin/streptomycin antibiotics (ThermoFisher Scientific, Waltham, MA, USA). All cells were incubated at 37 °C with 5% CO₂.