Purification and Ex Vivo Expansion of Fully Functional Salivary Gland Stem Cells

Hyposalivation often leads to irreversible and untreatable xerostomia. Salivary gland (SG) stem cell therapy is an attractive putative option to salvage these patients but is impeded by the limited availability of adult human tissue. Here, using murine SG cells, we demonstrate single-cell self-renewal, differentiation, enrichment of SG stem cells, and robust in vitro expansion. Dependent on stem cell marker expression, SG sphere-derived single cells could be differentiated in vitro into distinct lobular or ductal/lobular organoids, suggestive of progenitor or stem cell potency. Expanded cells were able to form miniglands/organoids containing multiple SG cell lineages. Expansion of these multipotent cells through serial passaging resulted in selection of a cell population, homogenous for stem cell marker expression (CD24hi/CD29hi). Cells highly expressing CD24 and CD29 could be prospectively isolated and were able to efficiently restore radiation-damaged SG function. Our approach will facilitate the use of adult SG stem cells for a variety of scientific and therapeutic purposes. •Single-cell-based in vitro self-renewal and differentiation of salivary gland stem cells•Functional rescue of hyposalivation with in vitro expanded salivary gland stem cells•Single-cell-derived organoids that contain salivary gland cell types Coppes and colleagues show single-cell CD24hi/CD29hi expansion, enrichment, and organoid differentiation of mouse salivary gland stem cells that potently rescue radiation-induced hyposalivation.