Fabrication protocol of tissue-engineered vocal fold mucosa.

<p>A 3 mm by 3 mm specimen of buccal mucosal tissue was excised. The mucosal section was divided into the epithelium and subepithelium. Isolated epithelial cells from the epithelium and isolated fibroblasts from the subepithelium were separately cultured for a two-week culture period. Following this, using cell-culture inserts, the proliferated fibroblasts were mixed into concentrated type I collagen gel on an oriented collagen sheet and cultured for one week. The proliferated epithelial cells were seeded and co-cultured on the collagen gel containing fibroblasts. After an additional two-week co-culture period, fabrication of the organotypic cultured tissue with stratified epithelial cells was complete.