Journal
JOURNAL OF CELLULAR PHYSIOLOGY
Volume 227, Issue 6, Pages 2654-2659Publisher
WILEY-BLACKWELL
DOI: 10.1002/jcp.23007
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Funding
- Pennsylvania Department of Health
- Pew Charitable Trust
- Landon AACR
- PhRMA Foundation
- American Cancer Society [117209-RSG-09-163-01-CNE]
- National Institutes of Health [F32 CA132450, P30 DK050306, R03 CA137811, P01 CA91955, P30 CA010815, R01 CA119224, R01 CA140657]
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Understanding the molecular and cellular processes underlying the development, maintenance, and progression of Barrett's esophagus (BE) presents an empirical challenge because there are no simple animal models and standard 2D cell culture can distort cellular processes. Here we describe a three-dimensional (3D) cell culture system to study BE. BE cell lines (CP-A, CP-B, CP-C, and CP-D) and esophageal squamous keratinocytes (EPC2) were cultured on a matrix consisting of esophageal fibroblasts and collagen. Comparison of growth and cytokeratin expression in the presence of all-trans retinoic acid or hydrochloric acid was made by immunohistochemistry and Alcian Blue staining to determine which treatments produced a BE phenotype of columnar cytokeratin expression in 3D culture. All-trans retinoic acid differentially affected the growth of BE cell lines in 3D culture. Notably, the non-dyplastic metaplasia-derived cell line (CP-A) expressed reduced squamous cytokeratins and enhanced columnar cytokeratins upon ATRA treatment. ATRA altered the EPC2 squamous cytokeratin profile towards a more columnar expression pattern. Cell lines derived from patients with high-grade dysplasia already expressed columnar cytokeratins and therefore did not show a systematic shift toward a more columnar phenotype with ATRA treatment. ATRA treatment, however, did reduce the squamoid-like multilayer stratification observed in all cell lines. As the first study to demonstrate long-term 3D growth of BE cell lines, we have determined that BE cells can be cultured for at least 3 weeks on a fibroblast/collagen matrix and that the use of ATRA causes a general reduction in squamous-like multilayered growth and an increase in columnar phenotype with the specific effects cell-line dependent. J. Cell. Physiol. 227: 26542659, 2012. (c) 2011 Wiley Periodicals, Inc.
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