Culturing Esophageal Cell Lines

Cell culture was similar to those we described previously[29 (link),32 (link),33 (link)]. Briefly, human esophageal squamous HET-1A cells were purchased from ATCC, Manassas, VA in 2011 and cultured in the bronchial epithelial cell medium (BEGM BulletKit, Cambrex, East Rutherford, NJ). Human Barrett’s cell line CP-A and Barrett’s dysplastic cell line CP-D were bought from ATCC (Manassas, VA) and cultured in Barrett’s medium containing keratinocyte medium-2 (Cambrex, Rockland, ME), 1.8 mmol/L CaCl₂, 5% fetal bovine serum, 400 ng/mL hydrocortisone, 20 ng/ml epidermal growth factor, 0.1 nmol/L cholera toxin, 20 μg/mL adenine, 5 μg/mL insulin, 70 μg/mL bovine pituitary extract, and antibiotics. EA cell line SK-GT-4 was purchased from Sigma and cultured in the Barrett’s medium.