Introducing a new type of organotypic laryngeal mucosa model

ABSTRACT: Research of human vocal fold (VF) biology is hampered by several factors. The sensitive microstructure of the VF mucosa is one of them and limits the in vivo research, as biopsies carry the unbearable risk of scarring. A laryngeal organotypic model consisting of VF epithelial cells and VF fibroblasts (VFF) might overcome some of the limitations. Whereas human VFF are available in several forms, availability of VF epithelial cells is scarce. Buccal mucosa might be a good source, as it is easily accessible, and biopsies heal without scarring. For this project we generated organotypic constructs consisting of immortalized human VF fibroblasts and primary human buccal epithelial cells. The constructs (n = 3) were compared to native laryngeal mucosa on a histological and proteomic level. The engineered constructs reassembled into a mucosa-like structure, after a cultivation period of 35 days. Immunohistochemical staining confirmed a multi-layered stratified epithelium, a collagen type IV positive barrier-like structure resembling the basal membrane, and an underlying layer containing VFF. Proteomic analysis revealed a total number of 1961 proteins. Of these, 83.8% were detected in both native VF and constructs, with only 53 proteins having significantly different abundance. 15.3% of detected proteins were identified in native VF mucosa only, most likely due to endothelial, immune and muscle cells within the VF samples, while 0.9% were found only in the constructs. Based on easily available cell sources, we could demonstrate that our organotypic laryngeal mucosa model shares many characteristics with native VF mucosa. It represents a stable and reproducible in vitro model and offers a wide range of possibilities ranging from the exploration of VF biology to the testing of interventions (e.g. drug testing).

INSTRUMENT(S): maXis

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Mucosa

SUBMITTER: Barbara Darnhofer  

LAB HEAD: Ruth Birner-Gruenberger

PROVIDER: PXD033688 | Pride | 2023-06-19

REPOSITORIES: Pride