Three-Dimensional Culture of Salivary Gland Stem Cell in Orthotropic Decellularized Extracellular Matrix Hydrogels

Radiotherapy in patients with cancer can kill cancer cells but also damage normal cells or tissues. During the treatment of patients with head and neck cancer or thyroid cancer, hyposalivation is a representative chronic side effect of radio-damaged salivary glands (SGs). The major symptom of hyposalivation is mouth dryness, resulting in several subsequent long-term complications. No effective therapeutic approaches have been developed to manage this symptom. In this study, we developed the first rat SG tissue-derived decellularized extracellular matrix hydrogel (DSGM-hydrogel) as a functional orthotropic bioscaffold for future efficient SG stem cell therapy. DSGM-hydrogels were characterized by rheological or biochemical analyses, and rat SG stem/progenitor cells (rSGSCs) were then subjected to three-dimensional culture in the DSGM-hydrogels. Interestingly, DSGM-hydrogel-embedded rSGSCs survived and expressed SG functional differentiation marker of amylase IA and increased enzyme activity of α-amylase in protein level, whereas they showed reduced levels of adult ductal stem/progenitor markers, including c-Kit, c-Met, and CD44. Furthermore, the expression levels of basic epithelial tight junction markers were recovered to levels similar to those naked SG tissues after culture in DSGM-hydrogels in transcription level. Therefore, our findings suggested that the DSGM-hydrogels could provide an appropriate microenvironment for stem/progenitor cell survival and a source of SG cytodifferentiation. This approach could be an applicable method to SG stem cell research as a potential source for an organoid and for clinical regenerative reagents to manage radio-damaged SGs in vivo. Impact Statement In this study, we established the first rat salivary gland (SG) tissue-derived decellularized extracellular matrix hydrogel (DSGM-hydrogel) and assessed the role of this hydrogel as a functional orthotropic bioscaffold. Our findings provide important insights into the applications of the DSGM-hydrogel as a biocompatible matrix for regenerative therapy of radio-damaged SGs.