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Title: | Reconstituted human upper airway epithelium as 3-d in vitro model for nasal polyposis. |
Author: | Callejas, Francisco de Borja Martínez Antón, Ma. Asunción Alobid, Isam Fuentes Prado, Mireya Cortijo, Julio Picado Vallés, César Roca i Ferrer, Jordi Mullol i Miret, Joaquim |
Keywords: | Cèl·lules epitelials Pòlips (Patologia) Diferenciació cel·lular Citoquines Inflamació Malalties del nas Epithelial cells Polyps (Pathology) Cell diferentiation Cytokines Inflammation Nose diseases |
Issue Date: | 19-Jun-2014 |
Publisher: | Public Library of Science (PLoS) |
Abstract: | BACKGROUND: Primary human airway epithelial cells cultured in an air-liquid interface (ALI) develop a well-differentiated epithelium. However, neither characterization of mucociliar differentiation overtime nor the inflammatory function of reconstituted nasal polyp (NP) epithelia have been described. OBJECTIVES: 1st) To develop and characterize the mucociliar differentiation overtime of human epithelial cells of chronic rhinosinusitis with nasal polyps (CRSwNP) in ALI culture system; 2nd) To corroborate that 3D in vitro model of NP reconstituted epithelium maintains, compared to control nasal mucosa (NM), an inflammatory function. METHODS: Epithelial cells were obtained from 9 NP and 7 control NM, and differentiated in ALI culture for 28 days. Mucociliary differentiation was characterized at different times (0, 7, 14, 21, and 28 days) using ultrastructure analysis by electron microscopy; ΔNp63 (basal stem/progenitor cell), β-tubulin IV (cilia), and MUC5AC (goblet cell) expression by immunocytochemistry; and mucous (MUC5AC, MUC5B) and serous (Lactoferrin) secretion by ELISA. Inflammatory function of ALI cultures (at days 0, 14, and 28) through cytokine (IL-8, IL-1β, IL-6, IL-10, TNF-α, and IL-12p70) and chemokine (RANTES, MIG, MCP-1, IP-10, eotaxin-1, and GM-CSF) production was analysed by CBA (Cytometric Bead Array). RESULTS: In both NP and control NM ALI cultures, pseudostratified epithelium with ciliated, mucus-secreting, and basal cells were observed by electron microscopy at days 14 and 28. Displaying epithelial cell re-differentation, β-tubulin IV and MUC5AC positive cells increased, while ΔNp63 positive cells decreased overtime. No significant differences were found overtime in MUC5AC, MUC5B, and lactoferrin secretions between both ALI cultures. IL-8 and GM-CSF were significantly increased in NP compared to control NM regenerated epithelia. CONCLUSION: Reconstituted epithelia from human NP epithelial cells cultured in ALI system provides a 3D in vitro model that could be useful both for studying the role of epithelium in CRSwNP while developing new therapeutic strategies, including cell therapy, for CRSwNP |
Note: | Reproducció del document publicat a: https://doi.org/10.1371/journal.pone.0100537 |
It is part of: | PLoS One, 2014, vol. 9, num. 6, p. e100537 |
URI: | http://hdl.handle.net/2445/122131 |
Related resource: | https://doi.org/10.1371/journal.pone.0100537 |
ISSN: | 1932-6203 |
Appears in Collections: | Articles publicats en revistes (IDIBAPS: Institut d'investigacions Biomèdiques August Pi i Sunyer) Articles publicats en revistes (Medicina) |
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