Formation Of Monolayers And Acinotubular Structures From Salivary Gland Cells

Objectives: To restore lost salivary epithelial function, either acinar cell renewal should be achieved or the function of the remnant ductal cells should be altered from an absorbing epithelium into a secretory epithelium. Our aim was to find optimal conditions to form functional epithelial monolayers and to from acinotubular structures using salivary cells originally isolated from rat or human salivary glands. Methods: Human cell cultures were prepared from dissected human submandibular glands. From culture day-2 either non-attaching supernatant cells (huSMG) or all surviving cells (PTHSG) were cultivated to obtain epithelial-like progenitor or mixed epithelial-mesenchymal progenitor culture, respectively. Passage 1-3 cells were seeded onto Transwell Clear permeable supports in either MEM or HepatoStim medium to prepare polarized monolayer. Par-C10 cells, originated from rat parotid gland, were cultured under standard conditions in DMEM/F12. Results: Autocrine factors and cell density are important for the growth and differentiation of huSMG and PTHSG cultures but not of Par-C10 cells. Both huSMG and PTHSG cultures grew to confluence as polarized monolayers on the permeable support in the presence of HepatoStim (TEER reached 700-1500 ohm*cm2) but not in MEM (TEER under 200 ohm*cm2). Culture in HepatoStim was required for the differentiation but not for the maintenance of human salivary gland epithelial cells. Acinar differentiation and acinotubular formation was succesfully achieved by basal membrane extract Matrigel in both Par-C10 and human salivary gland cultures. Conclusion: We have successfully established a functional model of human salivary gland epithelium and achieved acinar differentiation of rat and human salivary glands. Support:OTKA CK80928 and TÁMOP-4.2.1/B-09/1/KMR-2010-0001