Biosynthesis of a low-molecular-mass rat submandibular gland mucin glycoprotein in COS7 cells.
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ABSTRACT:
SUBMITTER: Nehrke K
PROVIDER: S-EPMC1218347 | biostudies-other | 1997 Apr
REPOSITORIES: biostudies-other
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Project description:PurposeHuman and rat salivary gland cell lines derived from tumors or genetic modification are currently available for research. Here, we attempted to culture and characterize long-term cultured cells spontaneously derived from wild type murine submandibular glands (SGs).MethodsSGs were removed from 3-week-old C57B/6J female mice and dissociated by collagenase type 1 and hyaluronidase digestion. Isolated SG epithelial cells were cultured in low calcium, serum-free growth media in the presence of cholera toxin (CT) during early passages. Single-cell colonies were isolated by limiting dilution culture after 25 passages. Early- and late-stage cell cultures were characterized for keratin 14, keratin 18, ?-smooth muscle actin, and p63 by immunostaining and quantitative real-time PCR analysis.ResultsSG epithelial cells cultured in optimized media maintained their proliferative ability and morphology for over 80 passages. Long-term cultured cells expressed keratin 14, keratin 18, and p63, indicative of an epithelial phenotype.ConclusionsEpithelial cells originating from wild type murine SGs could be cultured for longer periods of time and remain phenotypically similar to ductal basal epithelium.
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