Ultrastructural and immunohistochemical characterization of submandibular duct cells in culture and modification of outgrowth differentiation by manipulation of calcium ion concentration |
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Authors: | Barbara E Kurth Debra J Hazen-Martin Mary Ann Sens Donald A Sens |
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Institution: | (1) Department of Pathology, Medical University of South Carolina, 171 Ashley Avenue, 29425 Charleston, South Carolina |
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Abstract: | Summary The recognized need for epithelial cell culture models for cystic fibrosis (CF) research has resulted in ongoing efforts to
improve normal and CF submandibular duct cell culture capabilities. The duct is most likely the site of the CF defect in this
and other exocrine glands. In a previous report conditions required for the successful primary explant culture of normal and
CF submandibular glands were outlined; however, terminal keratinization and involution of these cultures were recognized as
severe limiting factors to their utilization in CF research. This report explores the effects of calcium concentrations in
the medium, growth factor supplements, and matrix components on growth and differentiation of these cultures. Results of the
study further confirm the ductal origin of cells in the outgrowth and demonstrate that progressive keratinization is initiated
only after cells proliferate beyond the environment of the explant fragment. Keratinization with subsequent multilayering,
desmosome formation, and involution in the cell outgrowth are governed in degree by the calcium concentration of the growth
medium. Upon reduction of medium calcium to 0.1 mM concentration, the cells proliferate as a monolayer and subculture through 8 to 9 passages and retain the capacity to undergo
ductlike differentiation.
This work was supported by Public Health Service grant AM 11028, Department of Health and Human Services, Washington, DC. |
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Keywords: | calcium ultrastructure immunohistochemistry submandibular duct differentiation |
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