Document Type
Peer-Reviewed Article
Publication Date
10-2019
Abstract
Objective: A current lack of methods for epithelial cell culture significantly hinders our understanding of the role of the epithelial and mucus barriers in vocal fold health and disease. Our first objective was to establish reproducible techniques for the isolation and culture of primary porcine vocal fold epithelial cells. Our second objective was to evaluate the functional significance of cell cultures using an in vitro exposure to an inflammatory cytokine. Methods: Epithelial cells were isolated from porcine vocal folds and expanded in culture. Characterization of cultures was completed by immunostaining with markers for pan-cytokeratin (epithelial cells), vimentin (stromal cells), von Willebrand factor (endothelial cell), and MUC1 and MUC4 (mucin) glycoproteins. Established epithelial cell cultures were then exposed to the inflammatory cytokine tumor necrosis factor alpha (TNF-α) for 24-hours, and transcript expression of MUC1 and MUC4 was evaluated. Results: Reproducible, porcine vocal fold epithelial cell cultures, demonstrating cobblestone appearance characteristic of the typical morphology of epithelial cell cultures were created. Cells showed positive staining for pan-cytokeratin with limited expression of vimentin and von Willebrand factor. Epithelial cells also expressed MUC1 and MUC4. TNF-α significantly increased transcript expression of MUC4. Conclusion: Here, we present the first report of successful culture of primary porcine vocal fold epithelial cells. Cultures will provide researchers with a valuable new in vitro tool to investigate vocal fold epithelium and mucus as well as the effects of common challenges, including inflammatory cytokines, on these barriers.
DOI
10.1002/lary.27909
PMID
30848488
Recommended Citation
Erickson-DiRenzo, Elizabeth; Leydon, Ciara; and Thibeault, Susan L., "Methodology for the Establishment of Primary Porcine Vocal Fold Epithelial Cell Cultures" (2019). Communication Disorders Faculty Publications. 178.
https://digitalcommons.sacredheart.edu/speech_fac/178
Publication
Laryngoscope
Volume
129
Issue
10
Publisher
Wiley
Pages
E355-E364
Comments
A portion of this work was presented at the 9th International Conference on Voice Physiology and Biomechanics, Salt Lake City, Utah, U.S.A., April 10, 2014. Ciara Leydon was then affiliated with the University of Wisconsin, Madison.
Version posted is the HHS Public Access Author Manuscript. Published in final edited form as: Laryngoscope. 2019 October ; 129(10): E355–E364. doi:10.1002/lary.27909.