Abstract
For alveolar type I cells, phenotype plasticity and physiology other than gas exchange await further clarification due to in vitro study difficulties in isolating and maintaining type I cells in primary culture. Using an established in vitro model of human fetal type II cells, in which the type II phenotype is induced and maintained by adding hormones, we assessed for transdifferentiation in culture toward a type I-like cell with hormone removal for up to 144 h, followed by electron microscopy, permeability studies, and RNA and protein analysis. Hormone withdrawal resulted in diminished type II cell characteristics, including decreased microvilli, lamellar bodies, and type II cell marker RNA and protein. There was a simultaneous increase in type I characteristics, including increased epithelial cell barrier function indicative of a tight monolayer and increased type I cell marker RNA and protein. Our results indicate that hormone removal from cultured human fetal type II cells results in transdifferentiation toward a type I-like cell. This model will be useful for continued in vitro studies of human fetal alveolar epithelial cell differentiation and phenotype plasticity.
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Abbreviations
- Cav-1:
-
caveolin-1
- DCI:
-
dexamethasone, cAMP, and isobutylmethylxanthine
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- PAI-1:
-
plasminogen activator inhibitor-1
- PGC:
-
pepsinogen C
- SEM:
-
scanning electron microscopy
- SP-A, SP-B, SP-C:
-
surfactant proteins A, B, C
- TEM:
-
transmission electron microscopy
- TER:
-
transepithelial resistance
- ZO-1:
-
zonula occludens-1
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Acknowledgements
The authors thank Sree Angampalli and Ping Wang for type II cell preparation, Michael Koval for assistance with the barrier studies, and Neelimah Shah at the Biomedical Imaging Core of the University of Pennsylvania School of Medicine for the microscopy images. They also thank Michael Beers and Harry Ischiropoulos for editorial assistance.
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NIH grants HL-077266 (C.D.F.) and HL059959 (S.H.G.) supported the research in this proposal.
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Foster, C., Varghese, L., Skalina, R. et al. In Vitro Transdifferentiation of Human Fetal Type II Cells Toward a Type I–like Cell. Pediatr Res 61, 404–409 (2007). https://doi.org/10.1203/pdr.0b013e3180332c6d
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DOI: https://doi.org/10.1203/pdr.0b013e3180332c6d
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