Abstract
Background
Bronchopulmonary dysplasia remains one of the most common complications of prematurity, despite significant improvements in perinatal care. Functional modeling of human lung development and disease, like BPD, is limited by our ability to access the lung and to maintain relevant progenitor cell populations in culture.
Methods
We supplemented Rho/SMAD signaling inhibition with mTOR inhibition to generate epithelial basal cell-like cell lines from tracheal aspirates of neonates.
Results
Single-cell RNA-sequencing confirmed the presence of epithelial cells in tracheal aspirates obtained from intubated neonates. Using Rho/SMAD/mTOR triple signaling inhibition, neonatal tracheal aspirate-derived (nTAD) basal cell-like cells can be expanded long term and retain the ability to differentiate into pseudostratified airway epithelium.
Conclusions
Our data demonstrate that neonatal tracheal aspirate-derived epithelial cells can provide a novel ex vivo human cellular model to study neonatal lung development and disease.
Impact
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Airway epithelial basal cell-like cell lines were derived from human neonatal tracheal aspirates.
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mTOR inhibition significantly extends in vitro proliferation of neonatal tracheal aspirate-derived basal cell-like cells (nTAD BCCs).
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nTAD BCCs can be differentiated into functional airway epithelium.
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nTAD BCCs provide a novel model to investigate perinatal lung development and diseases.
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Acknowledgements
This work was supported by the Department of Pediatrics, Massachusetts General Hospital. This work was also partly supported by the Harvard Stem Cell Institute (H.M.) and the Charles H. Hood Foundation (H.M.).
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J.L.: Substantial contributions to conception and design, acquisition of data, and analysis and interpretation of data; drafting the article or revising it critically for important intellectual content. X.Z.: Substantial contributions to conception and design, acquisition of data, and analysis and interpretation of data. J.E.S., L.X., T.G., C.Z., M.W., B.C.C.: Acquisition of data, and analysis and interpretation of data. J.C.L., H.L.: Analysis and interpretation of data. S.H.: Acquisition of data. H.M.: Substantial contributions to conception and design, analysis and interpretation of data; revising the article critically for important intellectual content. P.H.L.: Substantial contributions to conception and design, analysis and interpretation of data; revising the article critically for important intellectual content, and final approval of the version to be published.
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Patient samples were collected using informed consent forms approved by the Institutional Review Board of Partners HealthCare.
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Lu, J., Zhu, X., Shui, J.E. et al. Rho/SMAD/mTOR triple inhibition enables long-term expansion of human neonatal tracheal aspirate-derived airway basal cell-like cells. Pediatr Res 89, 502–509 (2021). https://doi.org/10.1038/s41390-020-0925-3
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DOI: https://doi.org/10.1038/s41390-020-0925-3
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