Derivation of Three-Dimensional Human Induced Pluripotent Stem Cell-Derived Vocal Fold Mucosa for Clinical and Pharmacological Applications

Abstract

Healthy vocal fold mucosa is composed of two major cell types, non-keratinized stratified squamous epithelium and vocal fold fibroblasts. Although dysfunction of the epithelium may play a significant pathogenic role in vocal fold diseases, studies at the genetic and molecular level using primary epithelial cells or models of human vocal fold mucosa have been significantly limited by the availability of relevant tissue types, poor growth, and heterogeneity of primary vocal fold epithelial cells. Here, we describe in vitro developmental differentiation of human induced pluripotent stem cells into vocal fold basal epithelial progenitors that were reseeded on collagen-fibroblast constructs to induce stratification and generate a three-dimensional model of human vocal fold mucosa. The engineered vocal fold mucosa represents physiologically relevant and clinically useful model that can be used as a tool for disease modeling and testing of therapeutic approaches for the treatment of laryngeal and VF inflammation.

Keywords: Developmental derivation; Medical research; Stem cells; Tissue engineering; Vocal fold mucosa.

Figure 1:

Summary of the differentiation protocol for the hiPSC-derived VF mucosa. Abbreviations: hiPSC, human induced pluripotent stem cells; AFE, Anterior Foregut Endoderm; A/Li, Air/Liquid Interface; DE, Definitive Endoderm; VBP, Vocal Fold Basal Progenitors; VFE, Vocal fold Epithelium.