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Differentiation potential of a basal epithelial cell line established from human bronchial explant

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Abstract

Due to the cellular complexity of the airway epithelium, it is important to carefully define bronchial cell lines that capture the phenotypic traits of a particular cell type. We describe the characterization of a human bronchial epithelial cell line, VA10. It was established by transfection of primary bronchial epithelial cells with retroviral constructs containing the E6 and E7 oncogenes from HPV16. The cell line has been cultured for over 2 yr, a total of 60 passages. Although prolonged culture resulted in increased chromosomal instability, no major phenotypic drift in marker expression was observed. The cells expressed cytokeratins 5, 13, 14, and 17 suggesting a basal-like phenotype. This was further supported by the expression of α6β4 integrins and the basal cell-associated transcription factor p63. The VA10 cell line generated high transepithelial electrical resistance in suspended and air–liquid interface culture, indicating functionally active tight junction (TJ) complexes. Immunocytochemistry showed the typical reticular structures of occludin and TJ-associated F-actin. VA10 produced pseudostratified layer in air–liquid interface culture with expression of p63 restricted to the basal layer. Furthermore, VA10 produced round colonies when cultured in laminin-rich reconstituted basement membrane, and immunostaining of claudin-1 and the basolateral marker β4 integrin revealed colonies that generated polarization as expected in vivo. These data indicate that VA10 epithelia have the potential to model the bronchial epithelium in vivo and may be useful to study epithelial regeneration and repair and the effect of chemicals and potential drug candidates on TJ molecules in airway epithelia.

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Acknowledgments

Grant support was provided by The Icelandic Research Council, Landspitali University Hospital Research Fund, University of Iceland Research Fund, and Science and Technology Policy Council-Thematic program in postgenomic biomedicine. We thank Prof. Michael J. Welsh at the University of Iowa for providing primary bronchial cells.

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Authors and Affiliations

  1. Biology Institute, University of Iceland, Reykjavik, Iceland

    Skarphedinn Halldorsson & Gudmundur Hrafn Gudmundsson

  2. Faculty of Pharmacy, University of Iceland, Reykjavik, Iceland

    Olafur Baldursson

  3. Faculty of Medicine, University of Iceland, Reykjavik, Iceland

    Valthor Asgrimsson, Ivar Axelsson & Thorarinn Gudjonsson

  4. Chromosome Laboratory, Department of Genetics and Molecular Medicine, Landspitali University Hospital, Reykjavik, Iceland

    Margret Steinarsdottir

  5. Division of Pulmonary Medicine, Landspitali University Hospital, Reykjavik, Iceland

    Olafur Baldursson

  6. Experimental Cell Biology Research Unit, Department of Hematology, Landspitali University Hospital, (K-building), 101, Reykjavik, Iceland

    Ivar Axelsson & Thorarinn Gudjonsson

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  1. Skarphedinn Halldorsson
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  2. Valthor Asgrimsson
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  3. Ivar Axelsson
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  4. Gudmundur Hrafn Gudmundsson
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  5. Margret Steinarsdottir
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  6. Olafur Baldursson
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  7. Thorarinn Gudjonsson
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Correspondence to Thorarinn Gudjonsson.

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Editor: J. Denry Sato

SH an VA contributed equally to this paper.

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Halldorsson, S., Asgrimsson, V., Axelsson, I. et al. Differentiation potential of a basal epithelial cell line established from human bronchial explant. In Vitro Cell.Dev.Biol.-Animal 43, 283–289 (2007). https://doi.org/10.1007/s11626-007-9050-4

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  • DOI: https://doi.org/10.1007/s11626-007-9050-4

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