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Sci Rep. 2017 Dec 21;7(1):17971. doi: 10.1038/s41598-017-17952-4.

Conditionally reprogrammed primary airway epithelial cells maintain morphology, lineage and disease specific functional characteristics.

Author information

1
Telethon Kids Institute, Centre for Health Research, The University of Western Australia, Crawley, Western Australia, Australia.
2
School of Paediatrics and Child Health, The University of Western Australia, Crawley, Western Australia, Australia.
3
Centre for Cell Therapy and Regenerative Medicine, School of Medicine and Pharmacology, The University of Western Australia, Nedlands, Western Australia, Australia.
4
Centre of Microscopy, Characterisation and Analysis, The University of Western Australia, Crawley, Western Australia, Australia.
5
School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, New South Wales, Australia.
6
Priority Research Centre for Asthma and Respiratory Disease, Hunter Medical Research Institute, Newcastle, New South Wales, Australia.
7
Department of Anesthesiology, Pharmacology and Therapeutics, University of British Columbia, Vancouver, Canada.
8
Telethon Kids Institute, Centre for Health Research, The University of Western Australia, Crawley, Western Australia, Australia. Anthony.Kicic@telethonkids.org.au.
9
School of Paediatrics and Child Health, The University of Western Australia, Crawley, Western Australia, Australia. Anthony.Kicic@telethonkids.org.au.
10
Centre for Cell Therapy and Regenerative Medicine, School of Medicine and Pharmacology, The University of Western Australia, Nedlands, Western Australia, Australia. Anthony.Kicic@telethonkids.org.au.
11
Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia. Anthony.Kicic@telethonkids.org.au.
12
Occupation and Environment, School of Public Health, Curtin University, Perth, Western Australia, Australia. Anthony.Kicic@telethonkids.org.au.
13
Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia.

Abstract

Current limitations to primary cell expansion led us to test whether airway epithelial cells derived from healthy children and those with asthma and cystic fibrosis (CF), co-cultured with an irradiated fibroblast feeder cell in F-medium containing 10 µM ROCK inhibitor could maintain their lineage during expansion and whether this is influenced by underlying disease status. Here, we show that conditionally reprogrammed airway epithelial cells (CRAECs) can be established from both healthy and diseased phenotypes. CRAECs can be expanded, cryopreserved and maintain phenotypes over at least 5 passages. Population doublings of CRAEC cultures were significantly greater than standard cultures, but maintained their lineage characteristics. CRAECs from all phenotypes were also capable of fully differentiating at air-liquid interface (ALI) and maintained disease specific characteristics including; defective CFTR channel function cultures and the inability to repair wounds. Our findings indicate that CRAECs derived from children maintain lineage, phenotypic and importantly disease-specific functional characteristics over a specified passage range.

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