Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
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Studies on the Air-Liquid Interface Culture as an Experimental Model for Physiology and Pharmacology of Tracheal Epithelial Cells

기관(氣管) 상피세포 생리 및 약리 실험모델로서의 공기-액체 접면 일차배양법 연구

  • 이충재 (충남대학교 의과대학 약리학 교실) ;
  • 이재흔 (충남대학교 의과대학 약리학 교실) ;
  • 석정호 (충남대학교 의과대학 약리학 교실) ;
  • 허강민 (충남대학교 의과대학 약리학 교실)
  • Published : 2002.12.01
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Abstract

In this study, we intended to get a preliminary data for establishing rat tracheal surface epithelial(RTSE) cell culture system as an experimental model for physiology and pharmacology of tracheal epithelial cells. Primary culture on the membrane support and application of the air-liquid interface system at the level of cell layer were performed. The cell growth rate and mucin production rate were measured according to the days in culture. The results were as follows: this culture system was found to manifest mucocilliary differentiation of rat tracheal epithelial cells, the cells were confluent and the quantity of produced and released mucin was highest on culture day 9, the mucin was mainly released to the apical side and tbe free $^3{H}$-glucosamine which was not incorporated to process of synthesis of mucin was left on the basolateral side. Taken together, we suggest that air-liquid interface culture system can be used as a substitute for immersion culture system and as an experimental model for in vivo mucus-hypersecretory diseases.

Keywords

References

  1. Adler, K.B., Schwarz, J.E., Anderson, W.H., and Welton, A.F. (1987). Platelet-activating factor stimulatcs secretion of mucin by cxplants of rodent airways in organ culture. Exp. Lung Res., 13, 25-43
  2. Chakrin, L.W., Baker, A.P., Spiser, S.S., Wardell, J.R.Jr., De Sanctis, N., and Dries, C. (1972). Am. Rev. Respir. Dis., 105, 368-381
  3. Clark, J.N. and Marchok, A.C. (1979). Characterization of mucin isolated from rat tracheal transplants. Biochim. Biophys. Acta.. 588, 357-367
  4. Coles, S.J., Neil, K.H, and Rcid, L.M. (1984). Potent stimulation of glycoprotein secretion in canine trachea by substance P. J. Appl. Physiol, 51, 1323-1327
  5. Gallagher, J.T. and Kent, P.W. (1975). Structure and metabolism of glycoproteins and glycosaminoglycans sccreted by organ cultures of rabbit trachea. Biochem. J., 148, 187-196 https://doi.org/10.1042/bj1480187
  6. Houdret, N., Lamblin, G., Scharfman, A., Humbert, P, and Roussel, P. (1983). Activation of bronchial mucin proteolysis by 4-aminophenylmercuric acetate and disulfide bond reducing agents. Biochim. Biophys. Acta., 758, 24-29
  7. Kaartinen L., Nettesheim, P., adler, K.B, and Randell, S.H. (1993). Rat tracheal cpithelial cell differentiation in vitro. In vitro Cell. Dev. Biol, 29A, 481-492
  8. Kim, K.C. and Brody, J.S. (1987). Gel contraction causcs mucin release in primary hamster tracheal epithelial cells growing on a collagcn gel. J. Cell. Biol., 105, 158a
  9. Kim, K.C. (1985). Possible requirement of collagen gel substraturn for production of mucinlike glycoproteins by primary rabbit tracheal epithelial cells in culture. In Vitro., 21, 617-621
  10. Kim, K.C., Opaskar-Hincman, H, and Bhaskar, K.R. (1989). Secretions from primary hamster tracheal surface epithelial cells in culture: Mucin-like glycoproteins, proteoglycans, and lipids. Exp. Lung Res., 15, 299-314
  11. Kim, K.C., Rearick, J.I, Nettesheim, P., and Jctten, A.M. (1985). Biochemical characterization of mucous glycoproteins synthesized and secreted by hamster tracheal epitlielial cells in primary culture. J. Biol. Chem., 260, 4021-4027
  12. Klinger, J.D., Tandler, B.T., Liedke, C.M., aiid Boat, T.F. (1984). Proteinases of Pseudomonas aeruginosa evoke mucin release by tracheal epithelium. J. Clin. Invest, 74, 1669-1678 https://doi.org/10.1172/JCI111583
  13. Ko, K.H. (1995). Inhibitor for secretion of mucin. The patent in thc Republic of Korea, Registered No. 6828
  14. Ko, K.H., Lee, C.J., Shin, C.Y., Jo, M.-J., and Kim, K.C. (1999). Inhibition of mucin release from airway goblet cells by polycationic peptides. Am. J. Physiol., 277(21), L811-L815
  15. Lechner, J.F., Haugen, A., Mcclendon, I.A., and Pettis, E.W. (1982). Clonal growth on normal adult human bronchial epithelial cells in a serum-free medium. In Vitro, 18, 800-812 https://doi.org/10.1007/BF02796504
  16. Marom, Z. Shelhamer, J.H., and Kaliner, M. (1981). Effects of arachidonic acid derivatives on the rclease of mucous glycoproteins from human airways. J. Clin. Invest., 67, 1695-1703 https://doi.org/10.1172/JCI110207
  17. Mian, N., Anderson, C.E., Pope, A.J., and Kent, P.W. (1982). Physicochemical properties of avian tracheal mucus. Biochem. J., 201, 533-542 https://doi.org/10.1042/bj2010533
  18. Niles, R.M., Christensen, T.G., Breuer, R., Stonc, P.J., and Snidcr, G.L. (1986). Serine proteases stimulate mucous glycoprotcin release from hamster tracheal ring organ culture. J. Lab. Clin. Med., 108, 489-497
  19. Rearick, J.L., Kim, K.C., Nettesheim, P., and Jetten, A.M. (1984). Hamster mucin secrcted in vitro contains poly-N-acetyllactosamine oligosaccharides. Fed. Proc., 43, 1696
  20. Shelhamer, J.H. Marom, Z., and Kaliner, M. (1980). Immunologic and neuropharmacologic stimulation of mucous glycoprotein release from human airways in vitro. J. Clin. Invest., 66, 1400-1408 https://doi.org/10.1172/JCI109993
  21. Shelhamer, J.H. Marom, Z., Logun, C., and Kaliner, M. (1984). Human respiratory mucous glycoproteins. Exp. Lung Res., 7, 149-162
  22. Sherman, J.M., Cheng, P.W., Tandler, B., and Boat, T.F. (1981). Mucous glycoproteins from cat tracheal goblet cells and mucous glands separated with EDTA. Am. Rev. Respir. Dis., 124, 476-479
  23. Wasano, K., Kim, K.C., Niles, R.M., and Brody, J.S. (1988). Membrane differcntiation markers of airway epithelial secretory cells. J. Histochem. Cytochem., 36, 167-178 https://doi.org/10.1177/36.2.3335774
  24. Woodward, H., Horscy, B., Bhavanandan, V.P. and Davidson, E.A. (1982). Isolation, Purification and properties of respiratory mucus glycoproteins. Biochemistry., 21, 694-701 https://doi.org/10.1021/bi00533a017
  25. Wu, R. and Smith, D. (1982) Continuous multiplication of rabbit tracheal epilhelial cclls in a defined, homone-sup-plemented medium. In Vitro, 18, 800-812 https://doi.org/10.1007/BF02796504