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Ultrastructural and autoradiographic observations of hamster cheek pouch epithelium grown in vitro
Authors:Baldev B Singh  George S Schuster  Virginia A Merchant  Virginia J Michelich
Institution:(1) Department of Oral Pathology, Medical College of Georgia, 30901 Augusta, Georgia;(2) Department of Oral Microbiology, Medical College of Georgia, 30901 Augusta, Georgia;(3) Department of Cell and Molecular Biology, Medical College of Georgia, 30901 Augusta, Georgia
Abstract:Summary Epithelial outgrowths from hamster cheek pouch explants were cultured for varying peroids of time up to 22 days. Growth of the epithelial sheets was monitored, employing colcemid for demonstrating mitotic activity and tritiated thymidine for DNA synthesis. Mitoses and thymidine uptake were observed among epithelial outgrowths at a considerable distance from the original explant. The epithelial nature of the growing cell sheets was confirmed, employing electron microscopic techniques. The cells exhibited the presence of tonofilaments, desmosomes, ribosomes, Golgi, mitochondria, and rough endoplasmic reticulum. The cultured explants were treated with cyclic nucleotides in order to investigate their modulatory effects on epithelial cell differentiation. Dibutyryl cAMP induced marked mitotic inhibition (46.3%) in our assay, which was increased to 57% with the addition of theophylline. Dibutyryl cGMP showed only a mild (5%) stimulatory effect on mitotic activity. Dibutyryl cAMP enhanced keratinization in the epithelial cell out-growths with the biogenesis of keratohyalin granules, whereas dibutyryl cGMP did not produce any observable alterations.
Keywords:cyclic nucleotides  epithelial cultures  ultrastructure  hamster cheek pouch
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