Development of a novel air-liquid interface airway tissue equivalent model for in vitro respiratory modeling studies.
Development of a novel air-liquid interface airway tissue equivalent model for in vitro respiratory modeling studies.
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DOI：
10.1038/s41598-023-36863-1
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发表时间：
2023-06-22
影响因子：
4.6
通讯作者：
Murphy, Sean V.
中科院分区：
文献类型：
--
作者：
Leach, Timothy;Gandhi, Uma;Reeves, Kimberly D.;Stumpf, Kristina;Okuda, Kenichi;Marini, Frank C.;Walker, Stephen J.;Boucher, Richard;Chan, Jeannie;Cox, Laura A.;Atala, Anthony;Murphy, Sean V.
人的呼吸道是复杂的结构，细胞、细胞外基质(ECM)蛋白和生物力学微环境之间存在着重要的相互作用。一个强大的、分化良好的体外培养系统能够准确地模拟这些相互作用，将为研究正常和病理的呼吸道生物学提供有用的工具。在这里，我们报道了一种生理相关的气液界面(ALI)三维呼吸道器官组织等效(OTE)模型的开发和表征，该模型具有三个新特征：天然肺成纤维细胞、溶解的肺ECM和具有可调节硬度和孔隙度的水凝胶基质。我们通过评估这些特征对人支气管上皮(HBE)细胞表型的影响，展示了OTE模型的多功能性。通过多光谱免疫组织化学和下一代测序评估上皮融合、跨上皮电阻值和上皮表型，分析该模型在ALI培养28天期间的变化。在水凝胶基质中同时包括溶解的肺ECM和天然肺成纤维细胞的培养形成分化良好的ALI培养物，该培养物保持屏障功能，并表达与杯状细胞、棒状细胞和纤毛细胞相关的成熟上皮标志物。水凝胶硬度的调节对HBE的分化没有负面影响，可以作为改变上皮表型的一个有价值的变量。这项研究强调了3D呼吸道OTE模型对人体呼吸道3D微环境的多个组成部分进行建模的可行性和通用性。
The human airways are complex structures with important interactions between cells, extracellular matrix (ECM) proteins and the biomechanical microenvironment. A robust, well-differentiated in vitro culture system that accurately models these interactions would provide a useful tool for studying normal and pathological airway biology. Here, we report the development and characterization of a physiologically relevant air–liquid interface (ALI) 3D airway ‘organ tissue equivalent’ (OTE) model with three novel features: native pulmonary fibroblasts, solubilized lung ECM, and hydrogel substrate with tunable stiffness and porosity. We demonstrate the versatility of the OTE model by evaluating the impact of these features on human bronchial epithelial (HBE) cell phenotype. Variations of this model were analyzed during 28 days of ALI culture by evaluating epithelial confluence, trans-epithelial electrical resistance, and epithelial phenotype via multispectral immuno-histochemistry and next-generation sequencing. Cultures that included both solubilized lung ECM and native pulmonary fibroblasts within the hydrogel substrate formed well-differentiated ALI cultures that maintained a barrier function and expressed mature epithelial markers relating to goblet, club, and ciliated cells. Modulation of hydrogel stiffness did not negatively impact HBE differentiation and could be a valuable variable to alter epithelial phenotype. This study highlights the feasibility and versatility of a 3D airway OTE model to model the multiple components of the human airway 3D microenvironment.
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影响因子：
3.3
作者：
Eskandari, Mona;Arvayo, Alberto L.;Levenston, Marc E.
通讯作者：
Levenston, Marc E.
DOI：
10.1165/rcmb.2009-0453oc
发表时间：
2011-04-01
影响因子：
6.4
作者：
Dvorak, Anna;Tilley, Ann E.;Crystal, Ronald G.
通讯作者：
Crystal, Ronald G.
DOI：
10.1016/j.ejpb.2012.12.014
发表时间：
2013-06-01
影响因子：
4.9
作者：
Blume, Cornelia;Davies, Donna E.
通讯作者：
Davies, Donna E.
DOI：
10.1513/pats.200801-010aw
发表时间：
2008-08-15
期刊：
Proceedings of the American Thoracic Society
影响因子：
--
作者：
Coraux, Christelle;Roux, Jacqueline;Birembaut, Philippe
通讯作者：
Birembaut, Philippe
影响因子：
4.6
作者：
De Rose V;Molloy K;Gohy S;Pilette C;Greene CM
通讯作者：
Greene CM