Isolation and Culture of Ferret Airway Stem Cells
Year: 2025
Type: article
Abstract: ) have gained considerable importance in respiratory disease research due to their notable susceptibility to these conditions and their anatomical similarities to humans. Here, we present a comprehensive description of the isolation and culture of stem/progenitor cells from the ferret airway, along with a protocol for their differentiation at the ALI. Our findings have demonstrated that this ferret culture system not only supports the differentiation of the predominant airway epithelial cell types but also facilitates the generation of rare airway epithelial subpopulations, including pulmonary ionocytes, tuft cells, and pulmonary neuroendocrine cells. Additionally, we provide a detailed procedure for measuring transepithelial ion transport relevant to airway diseases, particularly cystic fibrosis. The ability to isolate and culture ferret airway stem cells, combined with ALI differentiation and functional assessment of transepithelial ion transport, offers a powerful platform for evaluating genetic and pharmacologic interventions related to cystic fibrosis. Key features • A protocol for isolating ferret airway basal cells and generating air-liquid interface (ALI) cultures for electrophysiologic research. • Detailed procedures for propagating ferret airway basal cells and culturing in vitro well-differentiated airway epithelium. • A protocol for measuring ion transport, conductance, and immunofluorescence to identify airway cell types.
Source: BIO-PROTOCOL
Authors: Ziying Yan, John Engelhardt, Feng Yuan
Language: English
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: diamond
OpenAlex ID: W4412377992