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.
Keratinocyte medium 2
Manufactured by Cambrex
Keratinocyte medium-2 is a cell culture medium designed for the growth and maintenance of keratinocytes. It provides the necessary nutrients and growth factors to support the proliferation and differentiation of keratinocytes in vitro.
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Lab products found in correlation
2 protocols using keratinocyte medium 2
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Culturing Esophageal Cell Lines
2
Acid-Induced Changes in Barrett's Cells
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Human Barrett’s cell line BAR-T25 (link), provided to us by Dr. Rhonda Souza (University of Texas Southwestern Medical Center, Texas), was cultured in Keratinocyte Medium-2 (Ca2+-free solution, Cambrex, Rockland, ME) supplemented with 1.8 mM CaCl2 and other agents as we previously reported1 . For acid treatment, BAR-T cells were treated with acidic culture medium (pH 4.0) for 5 min, three times a day for 2, 4, 8 and 12 weeks. Cells were collected after acid treatment. To examine whether the molecular changes are reversible, cells were cultured at normal culture medium (pH 7.2) for additional 2, 4 and 8 weeks after acid treatment for 2, 4, 8 and 12 weeks, respectively.
BE EA cell line FLO-1 was generously provided to us by Dr. David Beer (University of Michigan). FLO cells were cultured in DMEM containing 10% fetal bovine serum and antibiotics. For PPI treatment, FLO-1 cells were treated with omeprazole 1μM for 24 hours.
BE EA cell line FLO-1 was generously provided to us by Dr. David Beer (University of Michigan). FLO cells were cultured in DMEM containing 10% fetal bovine serum and antibiotics. For PPI treatment, FLO-1 cells were treated with omeprazole 1μM for 24 hours.
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