THE PATHOBIOLOGY OF SALIVARY-GLAND .1. GROWTH AND DEVELOPMENT OF RAT SUBMANDIBULAR-GLAND ORGANOIDS CULTURED IN A COLLAGEN GEL MATRIX

Fragments of rat submandibular gland (organoids) which maintained the topological organization of the parent tissue were cultured in a three-dimensional collagen gel matrix for up to 30 days. At 48 h, vigorous peripheral outgrowth had occurred around each organoid. This was accompanied by central necrosis and the bridging of adjacent organoids. By day 5, large cyst-like spaces occupied the centre of many organoids. Bromodeoxyuridine labelling indicated that a considerable proportion of the lining cells were proliferating. Organoid growth peaked at between 5 and 10 days. Thereafter, the number of viable colonies and proliferating cells declined. Addition of isoproterenol after 24 h culture resulted in marked morphological alterations, with earlier and more prolific outgrowth and a greater tendency for organoids to flatten and grow out over the surface of the gel with squamous differentiation. Ultrastructurally, nuclear and cytoplasmic features of isoproterenol-treated and untreated cultures were similar. The secretory granules and extensive rough endoplasmic reticulum of terminal tubule cells, evident in organoids immediately after isolation, were infrequent after 24 h and absent by 48 h. Similar alterations occurred in the few acinar cells, so by 5 days the cultures were composed entirely of a uniform population of primitive, dedifferentiated cells. Further uses of this culture systems will include the study of diseases and disorders of the salivary glands as well as normal growth and differentiation pathways.