Abstract
Development of treatments for vocal dysphonia has been inhibited by lack of human vocal fold (VF) mucosa models because of difficulty in procuring VF epithelial cells, epithelial cells’ limited proliferative capacity and absence of cell lines. Here we report development of engineered VF mucosae from hiPSC, transfected via TALEN constructs for green fluorescent protein, that mimic development of VF epithelial cells in utero. Modulation of FGF signaling achieves stratified squamous epithelium from definitive and anterior foregut derived cultures. Robust culturing of these cells on collagen-fibroblast constructs produces three-dimensional models comparable to in vivo VF mucosa. Furthermore, we demonstrate mucosal inflammation upon exposure of these constructs to 5% cigarette smoke extract. Upregulation of pro-inflammatory genes in epithelium and fibroblasts leads to aberrant VF mucosa remodeling. Collectively, our results demonstrate that hiPSC-derived VF mucosa is a versatile tool for future investigation of genetic and molecular mechanisms underlying epithelium-fibroblasts interactions in health and disease.
Highlights
Development of treatments for vocal dysphonia has been inhibited by lack of human vocal fold (VF) mucosa models because of difficulty in procuring VF epithelial cells, epithelial cells’ limited proliferative capacity and absence of cell lines
We found a robust activation of pro-. In this investigation we developed an in vitro 3D model of human VF mucosa that provides a valuable model for investigation of VF epithelium and fibroblasts in health and disease
HiPSC-derived VF mucosae exhibited a slight variability in thickness, varying anywhere from 2 layers up to 10, and had variability in cell proliferation, as cell proliferation is likely correlated to thickness
Summary
Development of treatments for vocal dysphonia has been inhibited by lack of human vocal fold (VF) mucosa models because of difficulty in procuring VF epithelial cells, epithelial cells’ limited proliferative capacity and absence of cell lines. Modulation of FGF signaling achieves stratified squamous epithelium from definitive and anterior foregut derived cultures Robust culturing of these cells on collagen-fibroblast constructs produces three-dimensional models comparable to in vivo VF mucosa. Dysfunction of the epithelium may play a significant pathogenic role in VF diseases, studies at the genetic and molecular level using primary VF epithelial cells (VFEC) or models of human VF mucosa have been significantly limited by the availability of relevant tissue types derived from biopsies or cadaveric sources, poor growth, heterogeneity of primary VFEC, and absence of VFEC lines[13,14]. The main goals of this study were to generate reproducible hiPSC-derived
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
