Primaria plates and flasks

Immortalised BE cell lines BAR-T and CP-A (American Type Culture Collection, ATCC, Manassas, VA, United States) were cultured with epithelial cell medium 2 (ScienCell, Carlsbad, CA, United States), supplemented with 5% fetal bovine serum and antibiotics on primaria plates and flasks (BD Biosciences, Bedford, MA, United States). The immortalised human esophageal epithelia cell lines HET1A and EPC2 were obtained from ATCC. HET1A cell line was cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and antibiotics (Invitrogen). EPC2 cells were grown in keratinocyte SFM medium supplemented with 40 mg/mL bovine pituitary extract and 1.0 ng/mL epidermal growth factor (Invitrogen, Carlsbad, California, United States). All cell lines were grown at 37 °C in 5% carbon dioxide.

The immortalized human normal esophageal squamous cell line (HET1A) and the esophageal adenocarcinoma cell lines (FLO-1 and OE33) were obtained from American Type Culture Collection (ATCC, Manassas, VA) and were cultured in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% fetal bovine serum and antibiotics (Invitrogen, Carlsbad, CA). Immortalized Barrett's esophagus cell line (BAR-T, a kind gift from Dr. Rhonda Souza) was cultured with epithelial cell medium 2 (ScienCell, Carlsbad, CA), supplemented with 5% fetal bovine serum and antibiotics on primaria plates and flasks (BD Biosciences, Bedford, MA). All cell lines were grown at 37°C in 5% carbon dioxide.

DSC3 gene expression and DNA methylation were analyzed in 10 cell lines originating from esophagus, including normal esophageal squamous epithelium (HET1A and HEEC), Barrett's esophagus (BART, CPA, and CPB) and adenocarcinomas (OE33, FLO-1, OE19, SKGT4, and JHU-Eso-Ad1). These cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), Sigma Aldrich (St. Louis, MO, USA), ScienCell Research Laboratories (Carlsbad, CA, USA), Dr. Rhonda Souza (University of Texas Southwestern), Dr. David Beer (University of Michigan), and Dr. Jim Eshleman (John Hopkins University). The esophageal adenocarcinoma cell lines OE33, OE19, FLO-1, SKGT4, and JHU were cultured in Dulbecco's modified Eagle's medium (DMEM) media. The immortalized Barrett's esophagus cell lines were cultured with epithelial cell medium 2 (ScienCell). All the cell lines were supplemented with 10% fetal bovine serum and antibiotics (100 u/ml penicillin and 100 µg/ml streptomycin (Invitrogen, Carlsbad, CA, USA) on primaria plates and flasks (BD Biosciences, Bedford, MA, USA) in a 37°C incubator with 5% CO2.