Growth in primary culture of mouse submandibular epithelial cells embedded in collagen gels.

Mouse submandibular glands were dissociated and the epithelial cells embedded in a collagen gel matrix. A characteristic and reproducible pattern of growth was seen resulting in three-dimensional outgrowths with ductlike structures projecting into the matrix. A sustained cell growth leading to a 5 to 10-fold increase in cell number was observed in less than 2 wk. The extent of this growth was found to be dependent on serum concentration. Of the three sera tested, swine serum was found to promote greater growth compared to fetal bovine serum or horse serum. Swine serum dose response studies have shown that a concentration of 2 to 5% in the medium elicited only a modest increase, if any, in cell number compared to the initial value within a period of 2 wk. Various hormones and growth factors were then added to this "maintenance" medium. Insulin was found to stimulate growth consistently and reproducibly in a dose-dependent manner. Ultrastructurally, the resulting outgrowths were comprised of polarized cells joined by apical tight junction and desmosomes. The outgrowths produced epidermal growth factor in response to dihydrotestosterone, triiodothyronine, and cortisol. The present system provides a method for sustaining growth and functional differentiation in primary culture of mouse submandibular gland epithelial cells.