Three-dimensional cell culture for the study of nasal polyps

被引:1
作者
Franca, Carolina Nunes [1 ]
Lacerda Bachi, Andre Luis [1 ,2 ]
Kosugi, Eduardo Macoto [2 ]
Pezato, Rogerio [2 ,3 ]
Machado Santelli, Glaucia Maria [4 ]
do Amaral, Jonatas Bussador [2 ]
机构
[1] Univ Santo Amaro UNISA, Programa Posgrad Ciencias Saude, Sao Paulo, SP, Brazil
[2] Univ Fed Sao Paulo, Dept Otorrinolaringol & Cirurgia Cabeca & Pescoco, Lab Pesquisa ORL, Sao Paulo, SP, Brazil
[3] Univ Sao Paulo, Dept Otorrinolaringol & Oftalmol, Sao Paulo, SP, Brazil
[4] Univ Sao Paulo, Inst Ciencias Biomed, Dept Biol Celular & Desenvolvimento, Sao Paulo, SP, Brazil
关键词
3D cell culture; Spheroids; Chronic Rhinosinusitis with Nasal Polyps; Confocal microscopy; Morphology; CHRONIC RHINOSINUSITIS; MODEL; SPHEROIDS;
D O I
10.1016/j.bjorl.2021.11.001
中图分类号
R76 [耳鼻咽喉科学];
学科分类号
100213 ;
摘要
Objectives: Three-dimensional (3D) cell cultures have many applications such as stem cell biology research, new drug discovery, cancer, and Chronic Rhinosinusitis with Nasal Polyps (CRSwNP). This disease is characterized by a significant impact on quality of life and productivity. The diversity of factors that act in the progression of CRSwNP point to the creation of a cell culture model that allows the integration of different cell types with extracellular matrix. This work aimed to create a cell culture model in 3 dimensions (spheroids) for the study of Nasal Polyposis. Methods: Nasal polyp tissue from patients diagnosed with CRSwNP was mechanically dissociated using tweezers and a scalpel and the solution containing cells and small aggregates of nasal polyps was transferred to a Petri dish containing 5 mL of culture medium at the concentration of 10(6) cells/mL. Results: The spheroids were cultivated for 20 days, fixed and analyzed using confocal microscopy. In a 3D culture environment, the spheroids were formed both by clustering cells and from small tissue fragments. In the cultures analyzed, the ciliary beat was present from the dissociation of the cells up to 20 days in culture. Conclusion: Our findings also point to these characteristics showing the environment generated in our study, the cells remained differentiated for a longer time and with ciliary beating. Thus, this work shows that nasal polyp-derived cells can be maintained in a 3D environment, enabling better strategies for understanding CRSwNP in situations similar to those found in vivo. (c) 2021 Associacao Brasileira de Otorrinolaringologia e Cirurgia Cervico-Facial. Published by Elsevier Editora Ltda. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
引用
收藏
页码:S69 / S74
页数:6
相关论文
共 27 条
[1]  
Al-Sayed AA, 2017, LARYNGOSCOPE INVEST, V2, P398, DOI 10.1002/lio2.117
[2]   Guideline for the use of immunobiologicals in chronic rhinosinusitis with nasal polyps (RSCwPN) in Brazil [J].
Anselmo-Lima, Wilma T. ;
Tamashiro, Edwin ;
Romano, Fabrizio R. ;
Miyake, Marcel M. ;
Roithmann, Renato ;
Kosugi, Eduardo M. ;
Nakanishi, Marcio ;
Fornazieri, Marco A. ;
Bezerra, Thiago F. P. ;
Mello, Joao F. ;
Lessa, Marcus M. ;
Voegels, Richard L. ;
Piltcher, Otavio B. ;
Sakano, Eulalia ;
Valera, Fabiana C. P. .
BRAZILIAN JOURNAL OF OTORHINOLARYNGOLOGY, 2022, 88 (03) :471-480
[3]   Pathogenesis of chronic rhinosinusitis with nasal polyps: role of IL-6 in airway epithelial cell dysfunction [J].
Bequignon, Emilie ;
Mangin, David ;
Becaud, Justine ;
Pasquier, Jennifer ;
Angely, Christelle ;
Bottier, Mathieu ;
Escudier, Estelle ;
Isabey, Daniel ;
Filoche, Marcel ;
Louis, Bruno ;
Papon, Jean-Francois ;
Coste, Andre .
JOURNAL OF TRANSLATIONAL MEDICINE, 2020, 18 (01)
[4]  
Beule Achim G, 2010, GMS Curr Top Otorhinolaryngol Head Neck Surg, V9, pDoc07, DOI [10.1055/s-0029-1246124, 10.3205/cto000071]
[5]   CULTURED HUMAN NASAL EPITHELIAL MULTICELLULAR SPHEROIDS - POLAR CYST-LIKE MODEL TISSUES [J].
BRIDGES, MA ;
WALKER, DC ;
HARRIS, RA ;
WILSON, BR ;
DAVIDSON, AGF .
BIOCHEMISTRY AND CELL BIOLOGY-BIOCHIMIE ET BIOLOGIE CELLULAIRE, 1991, 69 (2-3) :102-108
[6]   Polarized expression of cystic fibrosis transmembrane conductance regulator and associated epithelial proteins during the regeneration of human airway surface epithelium in three-dimensional culture [J].
Castillon, N ;
Hinnrasky, J ;
Zahm, JM ;
Kaplan, H ;
Bonnet, N ;
Corlieu, P ;
Klossek, JM ;
Taouil, K ;
Avril-Delplanque, A ;
Péault, B ;
Puchelle, E .
LABORATORY INVESTIGATION, 2002, 82 (08) :989-998
[7]   Reconstituted Human Upper Airway Epithelium as 3-D In Vitro Model for Nasal Polyposis [J].
de Borja Callejas, Francisco ;
Martinez-Anton, Asuncion ;
Alobid, Isam ;
Fuentes, Mireya ;
Cortijo, Julio ;
Picado, Cesar ;
Roca-Ferrer, Jordi ;
Mullol, Joaquim .
PLOS ONE, 2014, 9 (06)
[8]   Alterations in cellular force parameters and cell projections in Nasal polyps-derived fibroblasts [J].
do Amaral, Jonatas Bussador ;
Bloise, Antonio Carlos ;
Franca, Carolina Nunes ;
Perez-Novo, Claudina ;
Machado-Santelli, Glaucia Maria ;
Alencar, Adriano Mesquita ;
Pezato, Rogerio .
AURIS NASUS LARYNX, 2020, 47 (01) :98-104
[9]  
do Amaral Jonatas Bussador, 2011, Naturalia (Rio Claro), V34, P1
[10]   MCF-7 Cells as a Three-Dimensional Model for the Study of Human Breast Cancer [J].
do Amaral, Jonatas Bussador ;
Rezende-Teixeira, Paula ;
Freitas, Vanessa Morais ;
Machado-Santelli, Glaucia Maria .
TISSUE ENGINEERING PART C-METHODS, 2011, 17 (11) :1097-1107