Summary
The growth, differentiation, ciliary beating pattern and frequency of human respiratory ciliated cells in primary culture were studied by scanning and transmission electron microscopy and by videomicroscopy. The epithelial cells were obtained as outgrowth from explants of adult nasal polyps. When the explants were grown on type-I and type-IV collagen substrates in a standard serum-free, hormone-supplemented medium, a high percentage of ciliated cells (range 29±5% to 37±6%) was present within 2 days of culture. After 5 days of culture, the percentage of ciliated cells near the explant was 51±5%. Most of the cultured ciliated cells (85%) were characterized by individual cilia showing a coordinated movement during the beat cycle and a beating frequency (13.3±1.3 Hz) similar to that reported in vivo. In the other 15% of the ciliated cells, the dyskinetic cilia were aggregated into clumps and characterized by a rigid and planar bending movement and a lower (P<0.01) beating frequency (10.7±1.4 Hz). It is suggested that the latter type of cell, already described during fetal development, might be an intermediate type of ciliated cell which appears temporarily during the surface respiratory epithelial differentiation.
Similar content being viewed by others
References
Bemis PA, Kennedy JR (1981) An improved model for studying the effect of Bordetella bronchiseptica on the ciliary activity of canine tracheal epithelial cells. J Infect Dis 144:349–357
Benali R, Dupuit F, Jacquot J, Fuchey C, Hinnrasky J, Ploton D, Puchelle E (1989) Growth and characterization of isolated bovine tracheal gland cells in culture. Influence of a reconstituted basement membrane matrix. Biol Cell 66:263–270
Boisvieux-Ulrich E, Laine MC, Sandoz D (1989) In vitro effects of taxol on ciliogenesis in quail oviduct. J Cell Sci 92:9–20
Chambard M, Gabrion J, Mauchamp J (1981) Influence of collagen gel on the orientation of epithelial cell polarity: follicle formation from isolated thyroid cells and performed monolayers. J Cell Biol 91:157–166
Chopra DP, Sullivan JK, Wille JJ, Siddiqui KM (1987) Propagation of differentiating normal human tracheobronchial epithelial cells in serum-free medium. J Cell Physiol 130:173–181
Chopra DP, Klinger MM, Sullivan JK (1989) Effect of vitamin A on growth and differentiation of human tracheobronchial epithelial cell cultures in serum-free medium. J Cell Sci 93:133–142
Crocker TT, O'Donnell TV, Nunes L (1973) Toxicity of benzo(a)pyrene and air pollution composite for adult human bronchial mucosa in organ culture. Cancer Res 33:88–93
Docherty RJ, Forrester JV, Lackie JM (1987) Type I collagen permits invasive behaviour by retinal pigmented epithelial cells in vitro. J Cell Sci 87:399–409
Gabridge MG, Gunderson H, Schaeffer SL, Barden-Stahl YD (1978) Ciliated respiratory epithelial monolayers; new model for Mycoplasma pneumoniae infection. Infect Immun 21:333–336
Gaillard D, Petit A, Lallement A, Puchelle E (1989) In vivo ciliogenesis in human fetal tracheal epithelium. Am J Anat 185:415–428
Kennedy JR, Ranyard JR (1983) Morphology and quantitation of ciliated outgrowths from cultured rabbit tracheal explants. Eur J Cell Biol 29:200–208
Lee TC, Wu R, Brody AR, Barrett JC, Nettesheim P (1984) Growth and differentiation of hamster tracheal epithelial cells in culture. Exp Lung Res 6:27–45
Opas M (1989) Expression of the differentiated phenotype by epithelial cells in vitro is regulated by both biochemistry and mechanics of the substratum. Dev Biol 131:281–293
Rossman CM, Forrest J, Newhouse MT (1980) Motile cilia in “immotile cilia” syndrome. Lancet I:1360
Sanderson MJ, Dirksen ER (1989) Mechanosensitive and beta-adrenergic control of the ciliary beat frequency of mammalian respiratory tract cells in culture. Am Rev Respir Dis 139:432–440
Sanderson MJ, Sleigh MA (1981) Ciliary activity of cultured rabbit tracheal epithelium: beat pattern and metachrony. J Cell Sci 47:331–347
Steele VE, Marchok AC, Nettesheim P (1978) Establishment of epithelial cell lines following exposure of cultured tracheal epithelium to 12-0-tetradecanolyphorbol-13-acetate. Cancer Res 38:3563–3567
Steinfort C, Wilson R, Mitchell T, Feldman C, Rutman A, Todd H, Sykes D, Walker J, Saunders K, Andrew PW, Boulnois GJ, Cole PJ (1989) Effect of Streptococcus pneumoniae on human respiratory epithelium in vitro. Infect Immun 57:2006–2013
Varsano S, Basbaum CB, Forsberg LS, Borson DB, Caughey G, Nadel JA (1987) Dog tracheal epithelial cells in culture synthesize sulfated macromolecular glycoconjugates and release them from the cell surface upon exposure to extracellular proteinases. Exp Lung Res 13:157–184
Whitcutt MJ, Adler KB, Wu R (1988) A biphasic chamber system for maintaining polarity of differentiation of cultured respiratory tract epithelial cells. In Vitro Cell Dev Biol 24:420–428
Wiesel JM, Gahiel H, Vlodavsky I, Gay I, Ben-Bassat H (1983) Cell attachment, growth characteristics and surface morphology of human upper-respiratory tract epithelium cultured on extracellular matrix. Eur J Clin Invest 13:57–63
Wu R (1986) In vitro differentiation of airway epithelial cells. In: Schiff LJ (ed) In vitro models of respiratory epithelium. CRC Press, Boca Raton Fla, pp 1–26
Wu R, Smith D (1982) Continuous multiplication of rabbit tracheal epithelial cells in a defined hormone-supplemented medium. In Vitro Cell Dev Biol 18:800–812
Wu R, Yankaskas J, Cheng E, Knowles MR, Boucher R (1985) Growth and differentiation of human nasal epithelial cells in culture: serum-free, hormone supplemented medium and proteoglycan synthesis. Am Rev Respir Dis 132:311–320
Yankaskas RJ, Cotton CU, Knowles MR, Gatzy JT, Boucher RC (1985) 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
Zahm JM, Dionisius JP, Pierrot D, Chevillard M, Puchelle E (1990) Mesure de la fréquence de battement ciliaire sur écran vidéo. Innov Tech Biol Med 11:118–129
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chevillard, M., Hinnrasky, J., Zahm, JM. et al. Proliferation, differentiation and ciliary beating of human respiratory ciliated cells in primary culture. Cell Tissue Res 264, 49–55 (1991). https://doi.org/10.1007/BF00305721
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00305721