Long-term culture of normal and cystic fibrosis epithelial cells grown under serum-free conditions Authors
Received: 11 September 1989 Accepted: 29 December 1989 DOI:
Cite this article as: Gruenert, D.C., Basbaum, C.B. & Widdicombe, J.H. In Vitro Cell Dev Biol (1990) 26: 411. doi:10.1007/BF02623833 Summary
The understanding of pathways associated with differentiated function in human epithelial cells has been enhanced by the development of methods for the short-term culture of human epithelial cells. In general these methods involve the use of serum. The subculture and maintenance of epithelial cells in long-term culture has been more problematic. A serum-free medium developed for human bronchial epithelial cells was slightly modified and found to be useful for the subculture and long-term maintenance of not only bronchial epithelial cells, but also tracheal, nasal polyp, and sweat gland epithelial cells from either normal or cystic fibrosis individuals. The cells maintained epithelial-specific characteristics after multiple subcultures. Monolayers of epithelial cells showed junctional complex formation, the presence of keratin, and micro villi. Functional studies with Ussing chambers showed short circuit current (I
sc) responses to isoproterenol, bradykinin, or calcium ionophore (A23187) in subcultured tracheal and bronchial cells. Key words human epithelial differentiation subculture cystic fibrosis
This work is supported by grants HL41928 and DK39619 (DCG), HL24136 (CBB), and HL42368 (JHW and DCG) from the National Institutes of Health, Bethesda, MD.
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