TY  - JOUR
TI  - Isolation and Culture of Ferret Airway Stem Cells
AU  - Yan, Ziying 
AU  - Engelhardt, John F.
AU  - Yuan, Feng 
VL  - 15
IS  - 14
PY  - 2025
DA  - 2025/07/20
SP  - e5399
C1  - Bio-protocol 2025;15:e5399
DO  - 10.21769/BioProtoc.5399
UR  - https://doi.org/10.21769/BioProtoc.5399
AB  - Well-differentiated airway epithelial cultures are commonly used to study airway stem cell lineages, ion and fluid transport, respiratory virus infection and replication, and disease mechanisms in vitro. This culture model involves the isolation and expansion of airway stem cells followed by their differentiation at an air–liquid interface (ALI), a process that has been previously documented in humans and mice. Domestic ferrets (Mustela putorius furo) 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.
KW  - Transgenic ferrets
KW  - Cystic fibrosis
KW  - Airway epithelium
KW  - CFTR
KW  - Ion transport
KW  - Air–liquid interface culture
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False