Summary
Bovine tracheal submucosal gland cells have been isolated by enzymatic digestion and serially propagated in tissue culture for more than 12 mo. (40 passages). The cells exhibit an epithelioid appearance at confluence and contain alcian blue (pH 2.5)/periodic acid-Schiff-positive material within cytoplasmic granules. By electron microscopy numerous osmiophilic secretory granules are seen. Maximal growth is observed when the cells are grown on human placental collagen-coated culture vessels in medium supplemented with 20% fetal bovine serum. Scintillation spectrometry revealed that radiolabeled precursor (35SO4) was incorporated into high molecular weight molecules and released from cells. Isoproterenol (10−6 to 10−3 M) stimulated the release of35SO4. The maximal response to isoproterenol was completely inhibited by the β-adrenergic antagonist propranolol. It is concluded that the cultured cells retain features of tracheal gland cells and may serve as a useful model of synthesis and secretion of macromolecules by tracheal gland cells.
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Barile, M. F.; Grabowski, M. W. Detection and identification of mycoplasmas in infected cell cultures by direct immunofluorescent staining. In: Tully, J. G.; Razin, S., eds. Methods in mycoplasmology, vol. 2. New York: Academic press; 1983:173–181.
Basbaum, C. B.; Mann, J.; Chow, A., et al. Monoclonal antibodies as probes for unique antigens in secretory cells of mixed exocrine organs. Proc. Natl. Acad. Sci. USA 81:4419–4423; 1984.
Basbaum, C. B.; Paul, A.; Finkbeiner, W. E., et al. Biochemical characterization of glycoconjugates secreted by bovine tracheal gland cells in culture (abstract). J. Cell Biol. 101:232a; 1985.
Beckstead, J. H. The evaluation of human lymph nodes, using plastic sections and enzyme histochemistry. Am. J. Clin. Pathol. 80:131; 1983.
Boersma, A.; Lamblin, G.; Degand, P., et al. Separation of a complex mixture of oligosaccharides by HPLC on bonded-primary amine packing using a linear-gradient solvent system. Carbohydr. Res. 94:C7-C9; 1981.
Bowes, D.; Corrin, B. Ultrastructural immunocytochemical localization of lysozyme in human bronchial glands. Thorax 32:163–170; 1977.
Chen, T. R. In situ demonstration of mycoplasma contamination in cell cultures by fluorescent Hoeschst 33258 stain. Exp. Cell Res. 104:255–262; 1977.
Coleman, D. L.; Tuet, I. K.; Widdicombe, J. H. Electrical properties of dog tracheal epithelial cells grown in monolayer culture. Am. J. Physiol. 246:C355-C359; 1984.
Daniel, P. F.; Wolf, G. Glycoprotein biosynthesis by organ cultures of hamster trachea. Biochim. Biophys. Acta 451:184–200; 1976.
Finkbeiner, W. E.; Basbaum, C. B. Monoclonal antibodies directed against human sputum: localization and periodate sensitivity of tracheal antigens (abstract). Am. Rev. Respir. Dis. 133:A77; 1986.
Finkbeiner, W. E.; Paul, A.; Basbaum, C. B. Isolation of bovine tracheal submucosal gland cells and establishment of a cell line (abstract), Fed. Proc. 44:642; 1985.
Goldman, W. E.; Baseman, J. B. Selective isolation and culture of a proliferating epithelial cell population from the hamster tracheal. In Vitro 16:313–319; 1980.
Hsu, T. C.; Benirschke, K. An atlas of mammalian chromosomes, vol. I. New York: Springer Verlag, Inc; 1967.
Jacquot, J.; Puchelle, E.; Basbaum, C., et al. Secretion of lysozyme by bovine tracheal serous cells in culture (abstract). Am. Rev. Respir. Dis. 133:A296; 1986.
Lamb, D.; Reid, L. Histochemical types of acidic glycoprotein produced by mucous cells of the tracheobraonchial glands in man. J. Pathol. 98:213–229; 1969.
Lamb, D.; Reid, L. Histochemical and autoradiographic investigation of the serous cells of the human bronchial glands. J. Pathol. 100:127–138; 1970.
Lamblin, G.; Boersma, A.; Klein, A., et al. Primary structure determination of 5 sialylated oligosaccharides derived from bronchial mucus glycoproteins of patients suffering from cystic fibrosis. J. Biol. Chem. 259:9051–9058; 1984.
Levy, J. A.; Summer, P. E.; Hooser, L. E. Rapid tissue culture method for detection of Mycoplasma hyorhinis. J. Gen. Microbiol. 128:2817–2820; 1982.
Lowry, O. H.; Rosebrough, N. J.; Farr, A. L., et al. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:265–275; 1951.
Mazzuca, M.; Lhermitte, M.; Lafitte, J.-J., et al. Use of lectins for detection of glycoconjugates in the glandular cells of the human bronchial mucosa. J. Histochem. Cytochem. 30:956–966; 1982.
Mossman, B. T.; Ezerman, E. B.; Alder, K. B., et al. Isolation and spontaneous transformation of cloned lines of hamster tracheal epithelial cells. Cancer Res. 40:4403–4409; 1980.
Phipps, R. J.; Williams, I. P.; Richardson, P. S., et al. Sympathomimetic drugs stimulate the output of secretory glycoproteins from human bronchi in vitro. Clin. Sci. 63:23–28; 1982.
Reid, L. Measurement of the bronchial mucous gland layer: a diagnostic yardstick in chronic bronchitis. Thorax 15:132–141; 1960.
Sachdev, G. P.; Fox, O. F.; Wen, G., et al. Isolation and characterization of glycoproteins from canine tracheal mucus. Biochim. Biophys. Acta 536:184–196; 1978.
Sherman, J. M.; Cheng, P. W.; Tandler, B., et al. Mucous glycoproteins from cat tracheal goblet cells and mucous glands separated with EDTA. Am. Rev. Respir. Dis. 124:476–479; 1981.
Slayter, H. S.; Lamblin, G.; Le Treut, A., et al. Complex structure of human bronchial mucus glycoprotein. Eur. J. Biochem. 142:209–218; 1984.
Spicer, S. S.; Mochizuki, I.; Setser, M. E., et al. Complex carbohydrates of rat tracheobronchial surface epithelium visualized ultrastructurally. Am. J. Anat. 158:93–109; 1980.
Thurlbeck, W. M.; Benjamin, B.; Reid, L. Development and distribution of mucous glands in the fetal human trachea. Br. J. Dis. Chest 550:49–53; 1961.
Tom-Moy, M.; Basbaum, C. B.; Nadel, J. A. Localization and release of lysozyme from ferret trachea: effects of adrenergic and cholinergic drugs. Cell Tissue Res. 228:549–562; 1983.
Tos, M. Development of the tracheal glands in man. Acta Pathol. Microbiol. Scand. 68[Suppl. 185]:1–130; 1966.
Worton, R. G.; Duff, C. Karyotyping. In: Jakoby, W. B.; Pastan, I. H., eds. New York: Academic Press; 1979:322–344.
Wu, R.; Smith, D. J. Continuous multiplication of rabbit tracheal epithelial cells in a defined, hormone-supplemented medium. In Vitro 18:800–812; 1982.
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This study was supported in part by NIH Program Project grant HL-24136, by a National Cystic Fibrosis Foundation Research Development Grant, and by a grant from Cystic Fibrosis Research, Inc. Dr. Finkbeiner is a recipient of NIH Clinical Investigator Award HL-01387.
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Finkbeiner, W.E., Nadel, J.A. & Basbaum, C.B. Establishment and characterization of a cell line derived from bovine tracheal glands. In Vitro Cell Dev Biol 22, 561–567 (1986). https://doi.org/10.1007/BF02623514
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DOI: https://doi.org/10.1007/BF02623514