Human alveolar epithelial cells expressing tight junctions to model the air-blood barrier

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Anna Kuehn
Stephanie Kletting
Cristiane de Souza Carvalho-Wodarz
Urska Repnik
Gareth Griffiths
Ulrike Fischer
Eckart Meese
Hanno Huwer
Dagmar Wirth
Tobias May
Nicole Schneider-Daum
Claus-Michael Lehr

Abstract

This paper describes a new human alveolar epithelial cell line (hAELVi – human Alveolar Epithelial Lentivirus immor­talized) with type I-like characteristics and functional tight junctions, suitable to model the air-blood barrier of the peripheral lung. Primary human alveolar epithelial cells were immortalized by a novel regimen, grown as monolayers on permeable filter supports and characterized morphologically, biochemically and biophysically. hAELVi cells maintain the capacity to form tight intercellular junctions, with high trans-epithelial electrical resistance (> 1000 Ω*cm²). The cells could be kept in culture over several days, up to passage 75, under liquid-liquid as well as air-liquid conditions. Ultra­structural analysis and real-time PCR revealed type I-like cell properties, such as the presence of caveolae, expression of caveolin-1, and absence of surfactant protein C. Accounting for the barrier properties, inter-digitations sealed with tight junctions and desmosomes were also observed. Low permeability of the hydrophilic marker sodium fluorescein confirmed the suitability of hAELVi cells for in vitro transport studies across the alveolar epithelium. These results suggest that hAELVi cells reflect the essential features of the air-blood barrier, as needed for an alternative to animal testing to study absorption and toxicity of inhaled drugs, chemicals and nanomaterials.

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How to Cite
[1]
Kuehn, A., Kletting, S., de Souza Carvalho-Wodarz, C., Repnik, U., Griffiths, G., Fischer, U., Meese, E., Huwer, H., Wirth, D., May, T., Schneider-Daum, N. and Lehr, C.-M. 2016. Human alveolar epithelial cells expressing tight junctions to model the air-blood barrier. ALTEX - Alternatives to animal experimentation. 33, 3 (Aug. 2016), 251-260. DOI:https://doi.org/10.14573/altex.1511131.
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