An in vitro airway wall model of remodeling

Choe, Melanie M.; Sporn, Peter H. S.

doi:10.1152/ajplung.00005.2003

Cited by 75 publications

(67 citation statements)

References 32 publications

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“…We first sought to optimize culture conditions for epithelial ciliogenesis. Compared with our previous work (11,12), we found that the presence of calcium, ethanolamine, and phospho-ethanolamine as well as higher concentrations of retinoic acid (30 ng/ml) and bovine pituitary extract (200 g/ml) improved the extent of ciliogenesis and cilia length (Fig. 3).…”

Section: Resultscontrasting

confidence: 63%

“…It is likely that this increased susceptibility is at least partly caused by epithelial activation and damage and by factors including cytokines secreted by mesenchymal and inflammatory cells, which in turn cause loss of barrier function. However, asthmatic airways also experience compressive strain due to frequent brochocon- striction, and it has been demonstrated in vitro that mechanical compressive stress itself, at physiologically relevant levels, can activate the epithelium in asthmatic-like ways and induce airway wall remodeling events in the absence of inflammatory mediators (11,12,52,60,67,68).…”

Section: Discussionmentioning

confidence: 99%

“…Coculture medium composition was varied to optimize ciliogenesis based on previously published protocols (12,45,55). The final optimized coculture medium, used for all the data presented in this study (medium A) consisted of 50% DMEM (low glucose, Invitrogen), 50% bronchial epithelial basal medium, 200 g/ml bovine pituitary extract (Pel-Freez Biologicals, Rogers, AR), 0.5 ng/ml human recombinant epidermal factor (Collaborative Research, Lexington, MA), 5 g/ml hydrocortisone, 0.5 g/ml epinepherine, 10 g/ml transferrin, 5 g/ml insulin, 30 ng/ml retinoic acid, 6.5 ng/ml triiodothyronine, 1.5 g/ml BSA, 12 g/ml calcium chloride, 30 ng/ml ethanolamine, 0.3 g/ml phosphoethanolamine, 50 g/ml gentamicin, and 50 ng/ml amphotericin-B (all from Sigma).…”

Section: Methodsmentioning

confidence: 99%

“…Protein dot blot was used to confirm mucin production as described previously (12). Briefly, samples and standards (mucin types I and III, Sigma) were filtered through a nitrocellulose membrane (NitroPure, GE Osmonics, Minnetonka, MN) using Bio-Dot SF apparatus (BioRad, Hercules, CA).…”

Section: Ajp-lung Cell Mol Physiolmentioning

confidence: 99%

“…In particular, several different in vitro models of the bronchial airway wall have been used to demonstrate that mechanical stress, at levels relevant to bronchoconstriction, can itself induce changes in gene and protein expression in epithelial cells and fibroblasts that are consistent with airway wall remodeling events, as well as matrix remodeling and smooth muscle cell (SMC) migration and proliferation (11,12,31,42,60,66,73).…”

mentioning

confidence: 99%

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Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model

Tomei¹,

Choe²

2008

American Journal of Physiology-Lung Cellular and Molecular Phys

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Tomei AA, Choe MM, Swartz MA. Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model. Am J Physiol Lung Cell Mol Physiol 294: L79-L86, 2008. First published November 16, 2007 doi:10.1152/ajplung.00062.2007.-Asthmatic patients are more susceptible to viral infection, and we asked whether dynamic strain on the airway wall (such as that associated with bronchoconstriction) would influence the rate of viral infection of the epithelial and subepithelial cells. To address this, we characterized the barrier function of a three-dimensional culture model of the bronchial airway wall mucosa, modified the culture conditions for optimization of ciliogenesis, and compared epithelial and subepithelial green fluorescent protein (GFP) transduction by a pWpts-GFP lentivirus, pseudotyped with VSV-G, under static vs. dynamic conditions. The model consisted of human lung fibroblasts, bronchial epithelial cells, and a type I collagen matrix, and after 21 days of culture at air liquid interface, it exhibited a pseudostratified epithelium comprised of basal cells, mucus-secreting cells, and ciliated columnar cells with beating cilia. Microparticle tracking revealed partial coordination of mucociliary transport among groups of cells. Slow dynamic compression of the airway wall model (15% strain at 0.1 Hz over 3 days) substantially enhanced GFP transduction of epithelial cells and underlying fibroblasts. Fibroblast-only controls showed a similar degree of transduction enhancement when undergoing dynamic strain, suggesting enhanced transport through the matrix. Tight junction loss in the epithelium after mechanical stress was observed by immunostaining. We conclude that dynamic compressive strain such as that associated with bronchoconstriction may promote transepithelial transport and enhance viral transgene delivery to epithelial and subepithelial cells. This finding has significance for asthma pathophysiology as well as for designing delivery strategies of viral gene therapies to the airways. mechanical stress; three-dimensional model; asthma; bronchoconstriction; ciliogenesis; human ASTHMA AND AIRWAY SUSCEPTIBILITY to viral infections have long been correlated clinically (2,9,16,26,36,61,62). Asthma is associated with recruitment of inflammatory cells, remodeling and thickening of the reticular basement membrane, subepithelial fibrosis, and airway smooth muscle hyperresponsiveness and hyperreactivity (reviewed in Refs. 7,17,19,21,23,25,48,70). The epithelial barrier function of asthmatic airways is often damaged (35), possibly increasing susceptibility to viral infection. Moreover, it has been shown that epithelial cells from asthmatic patients express a higher number of viral receptors on their surface (4, 62), which when activated exacerbate epithelial secretion of cytokines like granulocyte-macrophage colony-stimulating factor, interleukin (IL)-6, IL-8, IL-11, and RANTES that help maintain a persistent inflammatory state (46).Nevertheless, inflammatory mediators are only part of the story, as ...

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Section: Resultscontrasting

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Ajp-lung Cell Mol Physiolmentioning

confidence: 99%

mentioning

confidence: 99%

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Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model

Tomei¹,

Choe²

2008

American Journal of Physiology-Lung Cellular and Molecular Phys

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Cells and Culture Systems Used to Model the Small Airway Epithelium

Bhowmick

Gappa-Fahlenkamp

2016

Lung

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The pulmonary epithelium is divided into upper, lower, and alveolar (or small) airway epithelia and acts as the mechanical and immunological barrier between the external environment and the underlying submucosa. Of these, the small airway epithelium is the principal area of gas exchange and has high immunological activity, making it a major area of cell biology, immunology, and pharmaceutical research. As animal models do not faithfully represent the human pulmonary system and ex vivo human lung samples have reliability and availability issues, cell lines, and primary cells are widely used as small airway epithelial models. In vitro, these cells are mostly cultured as monolayers (2-dimensional cultures), either media submerged or at air-liquid interface. However, these 2-dimensional cultures lack a three dimension-a scaffolding extracellular matrix, which establishes the intercellular network in the in vivo airway epithelium. Therefore, 3-dimensional cell culture is currently a major area of development, where cells are cultured in a matrix or are cultured in a manner that they develop ECM-like scaffolds between them, thus mimicking the in vivo phenotype more faithfully. This review focuses on the commonly used small airway epithelial cells, their 2-dimensional and 3-dimensional culture techniques, and their comparative phenotype when cultured under these systems.

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Mechanobiology in the Third Dimension

2005

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Abstract-Cells are mechanically coupled to their extracellular environments, which play critical roles in both communicating the state of the mechanical environment to the cell as well as in mediating cellular response to a variety of stimuli. Along with the molecular composition and mechanical properties of the extracellular matrix (ECM), recent work has demonstrated the importance of dimensionality in cell-ECM associations for controlling the sensitive communication between cells and the ECM. Matrix forces are generally transmitted to cells differently when the cells are on two-dimensional (2D) vs. within three-dimensional (3D) matrices, and cells in 3D environments may experience mechanical signaling that is unique vis-à-vis cells in 2D environments, such as the recently described 3D-matrix adhesion assemblies. This review examines how the dimensionality of the extracellular environment can affect in vitro cell mechanobiology, focusing on collagen and fibrin systems.

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An in vitro airway wall model of remodeling

Cited by 75 publications

References 32 publications

Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model

Effects of dynamic compression on lentiviral transduction in an in vitro airway wall model

Cells and Culture Systems Used to Model the Small Airway Epithelium

Mechanobiology in the Third Dimension

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