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EBioMedicine. 2018 Jun 21. pii: S2352-3964(18)30197-X. doi: 10.1016/j.ebiom.2018.05.032. [Epub ahead of print]
Long-Term Culture of Distal Airway Epithelial Cells Allows Differentiation Towards Alveolar Epithelial Cells Suited for Influenza Virus Studies.
Imai-Matsushima A1, Martin-Sancho L1, Karlas A1, Imai S1, Zoranovic T1, Hocke AC2, Mollenkopf HJ3, Berger H1, Meyer TF4.
Author information
Abstract
As the target organ for numerous pathogens, the lung epithelium exerts critical functions in health and disease. However, research in this area has been hampered by the quiescence of the alveolar epithelium under standard culture conditions. Here, we used human distal airway epithelial cells (DAECs) to generate alveolar epithelial cells. Long-term, robust growth of human DAECs was achieved using co-culture with feeder cells and supplementation with epidermal growth factor (EGF), Rho-associated protein kinase inhibitor Y27632, and the Notch pathway inhibitor dibenzazepine (DBZ). Removal of feeders and priming with DBZ and a cocktail of lung maturation factors prevented the spontaneous differentiation into airway club cells and instead induced differentiation to alveolar epithelial cells. We successfully transferred this approach to chicken distal airway cells, thus generating a zoonotic infection model that enables studies on influenza A virus replication. These cells are also amenable for gene knockdown using RNAi technology, indicating the suitability of the model for mechanistic studies into lung function and disease.
KEYWORDS:
Alveolar epithelial cells; Distal airway epithelial cells; Influenza A virus; Lung disease
PMID: 29937069 DOI: 10.1016/j.ebiom.2018.05.032
Long-Term Culture of Distal Airway Epithelial Cells Allows Differentiation Towards Alveolar Epithelial Cells Suited for Influenza Virus Studies.
Imai-Matsushima A1, Martin-Sancho L1, Karlas A1, Imai S1, Zoranovic T1, Hocke AC2, Mollenkopf HJ3, Berger H1, Meyer TF4.
Author information
Abstract
As the target organ for numerous pathogens, the lung epithelium exerts critical functions in health and disease. However, research in this area has been hampered by the quiescence of the alveolar epithelium under standard culture conditions. Here, we used human distal airway epithelial cells (DAECs) to generate alveolar epithelial cells. Long-term, robust growth of human DAECs was achieved using co-culture with feeder cells and supplementation with epidermal growth factor (EGF), Rho-associated protein kinase inhibitor Y27632, and the Notch pathway inhibitor dibenzazepine (DBZ). Removal of feeders and priming with DBZ and a cocktail of lung maturation factors prevented the spontaneous differentiation into airway club cells and instead induced differentiation to alveolar epithelial cells. We successfully transferred this approach to chicken distal airway cells, thus generating a zoonotic infection model that enables studies on influenza A virus replication. These cells are also amenable for gene knockdown using RNAi technology, indicating the suitability of the model for mechanistic studies into lung function and disease.
KEYWORDS:
Alveolar epithelial cells; Distal airway epithelial cells; Influenza A virus; Lung disease
PMID: 29937069 DOI: 10.1016/j.ebiom.2018.05.032