Summary
We previously demonstrated that human tracheobronchial epithelial (TBE) cells synthesize mucin and form mucous granules in culture when they are maintained on a collagen gel (CG) substratum, but not on a plastic tissue culture surface or a thin collagen-coated surface (Wu et al., Am. J. Respir. Cell Mol. Biol., 3:467–478; 1990). This observation led us to examine the effects of CG thickness on cell growth and differentiation in primary human/monkey TBE cell cultures. Using the same CG preparation, culture dishes with different thicknesses of CG substratum were prepared. In general, equivalent degrees of cell attachment and proliferation were observed in all cultures maintained on a collagen gel, independent of the thicknesses of CG substratum. However, a greater degree of mucin synthesis and secretion by the cells was observed as the thickness of the CG substratum was increased. Cultures maintained on a thick collagen gel (1 mm) exhibited greater apical membrane complexity, more pseudostratification, and more mucous granules than did cultures maintained on a thin CG substratum. The optimal culture surface for airway mucous cell differentiation contains more than 1-mm thickness of collagen gel substratum.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Blau, H.; Guzouski, D. E.; Siddiqui, Z. A., et al. Fetal type 2 pneumocytes form alveolar-like structure and maintain long-term differentiation on extracellular matrix. J. Cell. Physiol. 136:203–214; 1989.
Burridge, K. Substrate adhesion in normal and transformed fibroblast: organization and regulation of cytoskeletal membrane and extracellular matrix components at focal contacts. Cancer Rev. 4:18–78; 1986.
Edelman, G. D. Cell adhesion molecules. Science 219:450–457; 1983.
Emerman, J. T.; Pitelka, D. R. Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes. In Vitro 13:316–328; 1977.
Fey, E. G.; Wan, K. M.; Penman, S. Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three dimensional organization and protein composition. J. Cell Biol. 98:1973–1984; 1984.
Horwitz, A.; Duggan, K.; Buck, C., et al. Interaction of plasma membrane fibronectin receptor with talin: a transmembrane linkage. Nature 320:531–533; 1986.
Juliano, R. L. Membrane receptors for extracellular matrix macromolecules: relationship to cell adhesion and tumor metatasis. Biochim. Biophys. Acta 907:261–278; 1987.
Lechner, J. F.; Stoner, G. D.; Yoakum, G. H., et al. In vitro carcinogenesis studies with human tracheobronchial tissues and cells. In: Schiff, L. J., ed. In vitro models of respiratory epithelium. Boca Raton, FL: CRC Press, Inc.; 1986:143–159.
Lechner, J. F.; Haugen, A.; McClendon, I. A., et al. Clonal growth of normal adult human bronchial epithelial cells in a serum-free medium. In Vitro 18:633–642; 1982.
Lee, T. C.; Wu, R.; Brody, A. R., et al. Growth and differentiation of hamster tracheal epithelial cells in culture. Exp. Lung Res. 6:27–45; 1984.
Lee, EY-H.; Lee, W. H.; Kaetzel, C. S., et al. Interaction of mouse mammary epithelial cells with collagen substrata: regulation of casein gene expression and secretion. Proc. Natl. Acad. Sci. USA 82:1419–1423; 1985.
Lin, H.; Carlson, D. M.; St. George, J. A., et al. An ELISA method for the quantitation of tracheal mucins from human and nonhuman primates. Am. J. Respir. Cell Mol. Biol. 1:41–48; 1989.
Martin, W. R.; Brown, C.; Zhang, Y-J., et al. Growth and differentiation of primary tracheal epithelial cells in culture: regulation by extracellular calcium. J. Cell. Physiol. 147:138–148; 1991.
Michalopoulos, G.; Pitot, H. C. Primary cultures of parenchymal liver cells on collagen membrane. Exp. Cell Res. 94:70–78; 1975.
Rearick, J. I.; Jetten, A. M. Effects of substratum and retinoids upon the mucosecretory differentiation of airway epithelial cells in vitro. Environ. Health Perspect. 80:229–237; 1989.
Robinson, C. B.; Wu, R. Culture of conducting airway epithelial cells in serum-free medium. J. Tissue Cult. Methods 13:95–102; 1991.
Ruoslahti, E.; Pierschbacher, M. D. New perspectives in cell adhesion: RGD and integrins. Science 238:491–497; 1987.
St. George, J. A.; Cranz, D. L.; Zicker, J. R., et al. An immunohistochemical characterization of rhesus monkey respiratory secretions using monoclonal antibodies. Am. Rev. Respir. Dis. 132:556–563; 1985.
Sporn, M. B.; Todaro, G. J. Autocrine secretion and malignant transformation of cells. N. Engl. J. Med. 303:878–880; 1980.
Van Scott, M. R.; Yankaskas, J. R.; Boucher, R. C. Culture of airway epithelial cells: research techniques. Exp. Lung Res. 11:75–94; 1986.
Wu, R.; Nolan, E.; Turner, C. Expression of tracheal differentiation functions in serum-free hormone-supplemented medium. J. Cell. Physiol. 125:167–181; 1985.
Wu, R. In vitro differentiation of airway epithelial cells. In: Schiff, L. J., ed. In vitro models of respiratory epithelium. Boca Raton, FL: CRC Press, Inc.; 1986:1–26.
Wu, R.; Martin, W. R.; Robinson, C. B., et al. Expression of mucin synthesis and secretion in human tracheobronchial epithelial cells grown in culture. Am. J. Respir. Cell Mol. Biol. 3:467–478; 1990.
Yang, J.; Nandi, S. Growth of cultured cells using collagen as substrate. Int. Rev. Cytol. 81:249–286; 1983.
Yankaskas, J. R.; Cotton, C. U.; Knowles, M. R., et al. Culture of human nasal epithelial cells on collagen matrix supports: a comparison of bioelectric properties of normal and cystic fibrosis epithelia. Am. Rev. Respir. Dis 132:1281–1287; 1985.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Robinson, C.B., Wu, R. Mucin synthesis and secretion by cultured tracheal cells: effects of collagen gel substratum thickness. In Vitro Cell Dev Biol - Animal 29, 469–477 (1993). https://doi.org/10.1007/BF02639381
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02639381