Journal article
Development and characterization of ferret ex vivo tracheal injury and cell engraftment model
Frontiers in medicine, Vol.10, 1144754
04/01/2023
DOI: 10.3389/fmed.2023.1144754
PMCID: PMC10126424
PMID: 37113613
Abstract
The field of airway biology research relies primarily on in vitro and in vivo models of disease and injury. The use of ex vivo models to study airway injury and cell-based therapies remains largely unexplored although such models have the potential to overcome certain limitations of working with live animals and may more closely replicate in vivo processes than in vitro models can. Here, we characterized a ferret ex vivo tracheal injury and cell engraftment model. We describe a protocol for whole-mount staining of cleared tracheal explants, and showed that it provides a more comprehensive structural overview of the surface airway epithelium (SAE) and submucosal glands (SMGs) than 2D sections, revealing previously underappreciated structural anatomy of tracheal innervation and vascularization. Using an ex vivo model of tracheal injury, we evaluated the injury responses in the SAE and SMGs that turned out to be consistent with published in vivo work. We used this model to assess factors that influence engraftment of transgenic cells, providing a system for optimizing cell-based therapies. Finally, we developed a novel 3D-printed reusable culture chamber that enables live imaging of tracheal explants and differentiation of engrafted cells at an air-liquid interface. These approaches promise to be useful for modeling pulmonary diseases and testing therapies.Graphical abstract1,2. We describe here a method for differential mechanical injury of ferret tracheal explants that can be used to evaluate airway injury responses ex vivo. 3. Injured explants can be cultured at ALI (using the novel tissue-transwell device on the right) and submerged long-term to evaluate tissue-autonomous regeneration responses. 4. Tracheal explants can also be used for low throughput screens of compounds to improve cell engraftment efficiency or can be seeded with particular cells to model a disease phenotype. 5. Lastly, we demonstrate that ex vivo-cultured tracheal explants can be evaluated by various molecular assays and by immunofluorescent imaging that can be performed live using our custom-designed tissue-transwell.
Details
- Title: Subtitle
- Development and characterization of ferret ex vivo tracheal injury and cell engraftment model
- Creators
- Vitaly Ievlev - University of IowaAlbert C. Pai - University of IowaDrew S. Dillon - Protostudios, Carver College of Medicine, University of Iowa, Iowa City, IA, United StatesSpencer Kuhl - Protostudios, Carver College of Medicine, University of Iowa, Iowa City, IA, United StatesThomas J. Lynch - University of IowaKyle W. Freischlag - Department of Cardiothoracic Surgery, Carver College of Medicine, University of Iowa Hospitals and Clinics, Iowa City, IA, United StatesCaitlyn B. Gries - University of IowaJohn F. Engelhardt - University of IowaKalpaj R. Parekh - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Frontiers in medicine, Vol.10, 1144754
- DOI
- 10.3389/fmed.2023.1144754
- PMID
- 37113613
- PMCID
- PMC10126424
- NLM abbreviation
- Front Med (Lausanne)
- eISSN
- 2296-858X
- Publisher
- Frontiers Media S.A
- Language
- English
- Date published
- 04/01/2023
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Cardiothoracic Surgery; Internal Medicine
- Record Identifier
- 9984398346902771
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