Cultures of human tracheal gland cells of mucous or serous phenotype

• Published in: In vitro cellular & developmental biology. Animal 2010-05, Vol.46 (5), p.450-456
• Main Authors: Finkbeiner, Walter E, Zlock, Lorna T, Mehdi, Irum, Widdicombe, Jonathan H
• Format: Article
• Language: English
• Subjects: Animal Genetics and Genomics; Antimicrobial agents; Biomedical and Life Sciences; CELL AND TISSUE MODELS; Cell Biology; Cell Culture; Cell culture techniques; Cell growth; Cells, Cultured; Culture Media; Cultured cells; Developmental Biology; Epidermal growth factor; Epithelial cells; Epithelium; Exocrine Glands - cytology; Filters; Glands; Granules; Humans; lactoferrin; Lactoferrin - metabolism; Life Sciences; Liquids; Lysozyme; Microscopy, Electron; mucin; Mucins; Phenotype; Phenotypes; polycarbonate; polyethylene terephthalate; Respiratory Mucosa - cytology; Respiratory tract; Secretion; Serous Membrane - cytology; Short-circuit current; Stem Cells; Tight junctions; Trachea - cytology; Tubules; Ultrastructure
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-009-9262-x

There are two main epithelial cell types in the secretory tubules of mammalian glands: serous and mucous. The former is believed to secrete predominantly water and antimicrobials, the latter mucins. Primary cultures of human airway gland epithelium have been available for almost 20 yr, but they are poorly differentiated and lack clear features of either serous or mucous cells. In this study, by varying growth supports and media, we have produced cultures from human airway glands that in terms of their ultrastructure and secretory products resemble either mucous or serous cells. Of four types of porous-bottomed insert tested, polycarbonate filters (Transwells) most strongly promoted the mucous phenotype. Coupled with the addition of epidermal growth factor (EGF), this growth support produced “mucous” cells that contained the large electron-lucent granules characteristic of native mucous cells, but lacked the small electron-dense granules characteristic of serous cells. Furthermore, they showed high levels of mucin secretion and low levels of release of lactoferrin and lysozyme (markers of native serous cells). By contrast, growth on polyethylene terephthalate filters (Cyclopore) in medium lacking EGF produced “serous” cells in which small electron-dense granules replaced the electron-lucent ones, and the cells had high levels of lactoferrin and lysozyme but low levels of mucins. Measurements of transepithelial resistance and short-circuit current showed that both “serous” and “mucous” cell cultures possessed tight junctions, had become polarized, and were actively secreting Cl.