Dye-coupling in three-dimensional histoculture of rat lingual frenulum |
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Authors: | Leonid Margolis Boris Baibakov Carlos Collin Sidney A. Simon |
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Affiliation: | (1) Laboratory of Theoretical and Physical Biology, National Institutes of Health, Building 10, Room 10D14, 20892 Bethesda, Maryland;(2) Belozorsky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia;(3) Departments of Neurobiology and Anesthesiology, Duke University, 27710 Durham, North Carolina |
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Abstract: | Summary A three-dimensional histoculture of wet stratified squamous epithelium of rat lingual frenulum was cultured on a liquid-air interface. The tissue retained its morphology for many days in culture. During this period the vast majority of the epithelial cells remained viable and exhibited dye (lucifer yellow) coupling in all living epithelial strata. Dye coupling was determined using two methods: the conventional intracellular injection method, and a new method—“cut-loading.” In the cut-loading method, an incision is made in the epithelium in the presence of dye, and intracellular diffusion of dye throughout the epithelium was measured using confocal microscopy. The basolateral surface of the lingual frenulum also acted as a substrate for neuroblastoma cells to grow without exogenously added trophic factors. These neuroblastoma cells grow neurites that establish contacts with epithelial cells. This preparation can serve as a model for investigating interactions among epithelial cells and between nerves and epithelial cells. |
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Keywords: | gap junctions tongue stratified epithelium tissue culture |
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