Growth in primary culture of mouse submandibular epithelial cells embedded in collagen gels |
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Authors: | Jason Yang Darragh Flynn Lisa Larson Susan Hamamoto |
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Institution: | (1) Cancer Research Laboratory, University of California, 230 Warren Hall, 94720 Berkeley, California |
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Abstract: | Summary 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 junctions and desmosomes. These 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.
This investigation was supported by PHS Grants CA05388 and CA09041, awarded by the National Cancer Institute, Department of
Health and Human Services. |
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Keywords: | growth primary culture submandibular gland collagen |
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