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

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Anna Kuehn 1, Stephanie Kletting 1, Cristiane de Souza Carvalho-Wodarz 1, Urska Repnik 3, Gareth Griffiths 3, Ulrike Fischer 4, Eckart Meese 4, Hanno Huwer 5, Dagmar Wirth 6,8, Tobias May 7, Nicole Schneider-Daum 1 and Claus-Michael Lehr 1,2
1 Drug Delivery (DDEL), Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Saarbrücken, Germany
2 Department of Pharmacy, Saarland University, Saarbrücken, Germany
3 Department of Biosciences, University of Oslo, Oslo, Norway
4 Saarland University, Institute of Human Genetics, University Hospital, Homburg/Saar, Germany
5 SHG Clinics, Department of Cardiothoracic Surgery, Völklingen Heart Center, Völklingen, Germany
6 Model Systems for Infection and Immunity, Helmholtz-Centre for Infection Research (HZI), Braunschweig, Germany
7 InSCREENeX GmbH, Braunschweig, Germany; 8Experimental Hematology, Medical School Hannover, Hannover, Germany


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.


Keywords: lentivirus immortalization, ATI cells, pulmonary drug delivery, inhalation toxicology


ALTEX 33(3), 251-260

doi: 10.14573/altex.1511131

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