Abstract
The tongue is a complex organ involved in a variety of functions such as mastication, speech, and taste sensory function. Enzymatic digestion techniques have been developed to allow the dissociation of the epithelium from the connective tissue of the tongue. However, it is not clear if the integrity and three-dimensional architecture of the isolated epithelium is preserved, and, furthermore if this tissue separation technique excludes its contamination from the mesenchymal tissue. Here, we first describe in detail the methodology of tongue epithelium isolation, and thereafter we analyzed the multicellular compartmentalization of the epithelium by three-dimensional fluorescent imaging and quantitative real-time PCR. Molecular characterization at both protein and transcript levels confirmed the exclusive expression of epithelial markers in the isolated epithelial compartment of the tongue. Confocal imaging analysis revealed that the integrity of the epithelium was not affected, even in the basal layer, where areas of active cell proliferations were detected. Therefore, the preservation of both the architecture and the molecular signature of the tongue epithelium upon enzymatic tissue separation enable further cellular, molecular and imaging studies on the physiology, pathology, and regeneration of the tongue.
Highlights
The tongue is a complex organ located on the floor of the mouth whose muscular core is protected by a multi-layered epithelial mucosa
Advanced confocal imaging was used and we observed that the three-dimensional (3D) structure of the epithelium remained unaffected after the tissue separation procedure
Enzymatic tissue separation procedures have previously been applied in regenerative approaches using 3D tongue epithelium and taste bud organoids derived from isolated epithelial stem cells (Ren et al, 2014; Aihara et al, 2015; Hisha et al, 2016; Hisha and Ueno, 2016; Palechor-Ceron et al, 2019)
Summary
The tongue is a complex organ located on the floor of the mouth whose muscular core is protected by a multi-layered epithelial mucosa. While the ventral part of the tongue is lined by a thin nonkeratinized epithelium, its dorsal part is covered by a specialized, stratified, highly keratinized mucosa that consists of four types of papillae (Takahashi et al, 2019). The basal layer of the oral epithelium and the underlying mesenchyme are separated by a basement membrane called the lamina propria (Squier and Brogden, 2011; Castillo-Azofeifa et al, 2018). The generated undifferentiated progenitors translocate to the epithelial layer and, after progressing through the amplification stage, start to differentiate and progressively acquire elongated cell shapes (Watt, 1989; Castillo-Azofeifa et al, 2018). The study of the epithelial stem cells of the tongue has been hampered by the lack of efficient tools for the visualization and manipulation of the deeper epithelial layers, such as the basal layer
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
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 call
