A NOVEL MAGNETIC THREE-DIMENSIONAL LEVITATED PRIMARY CELL CULTURE SYSTEM FOR THE DEVELOPMENT OF IN VITRO SALIVARY GLAND-LIKE ORGANOIDS

Abstract

Salivary gland (SG) hypofunction and dry mouth leads to oral complications and impaired quality of life. Recently, researchers have proposed to repair hypo-functional SG via various cell-based approaches in three-dimensional (3D) scaffold-based systems. The majority have nonhuman-based components. Therefore, this project aims to develop SG mini-glands/organoids using a novel 3D xeno-free system (M3DL) that assembles and levitates magnetized primary SG-derived cells (SGDC). To accomplish this, SGDC were isolated, expanded, and characterized by flow cytometry, qPCR and immunofluorescence imaging to assess SG-specific markers. Further, SGDC were assembled in M3DL to generate SG-like 3D organoids for 7 days. The resulting organoids presented increased biocompatibility compared to conventional salispheres in cell viability assays. Moreover, the organoids expressed SG markers from different cellular compartments (acinar/ductal epithelial, myoepithelial and neuronal). Intracellular calcium and α-amylase activity assays showed functional organoids with SG-specific secretory activity. Thus, these outcomes suggest that M3DL is a promising tool to generate SG-like organoids for SG repair.