Three-dimensional organotypic culture of human salivary glands: the slice culture model

ObjectiveA challenge in studying human salivary glands is to maintain the cells exvivo in their three-dimensional (3D) morphology with an intact native extracellular matrix (ECM) environment. This paper established a human salivary 3D organotypic slice culture model that could maintain its physiological functions as well as allowing a direct visualization of the cells. MethodsHuman salivary biopsies from six patients were embedded in agarose and submerged in cold buffer for thin (50m) sectioning using a vibratome. Salivary slices' were mechanically supported by a porous membrane insert that allowed an air-liquid interface and cultured in serum-free culture media. Cell viability, proliferation, apoptosis, physiological functions, and gene expression were assessed during 14days of culture. ResultsHuman salivary slices maintained cell survival (70-40%) and proliferation (6-17%) for 14days exvivo. The protein secretory (amylase) function decreased, but fluid (intracellular calcium mobilization) function was maintained. Acinar, ductal, and myoepithelial cell populations survived and maintained their 3D organization within the slice culture model. ConclusionThe human salivary slice culture model kept cells alive exvivo for 14days as well as maintaining their 3D morphology and physiological functions.