Epithelial cell medium 2

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.

Three ESCC cell lines (KYSE150, KYSE30, and EC109) were obtained from Shanghai Institutes for Biological Sciences (Shanghai, China). They were cultured in RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) in a humidified atmosphere of 5% CO₂ at 37°C. HEsEpiC esophageal epithelial cells were purchased from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured in epithelial cell medium-2 (ScienCell Research Laboratories).