Murine Alveolar Epithelial Cells and Their Lentivirus-mediated Immortalisation. Issue 2 (May 2018)

• Record Type: Journal Article
• Authors: Sapich, Sandra; Hittinger, Marius; Hendrix-Jastrzebski, Remi; Repnik, Urska; Griffiths, Gareth; May, Tobias; Wirth, Dagmar; Bals, Robert; Schneider-Daum, Nicole; Lehr, Claus-Michael
• Is Part Of: ATLA. Volume 46:Issue 2(2018)
• Page Start: 73
• Page End: 89
• Publication Date: 2018-05
• Subjects: cytotoxicity -- mouse cell line -- pulmonary drug delivery -- respiratory in vitro model -- Three Rs; Laboratory animals -- Periodicals; 616.0273
• DOI: 10.1177/026119291804600207
• Languages: English
• ISSNs: 0261-1929

Abstract

In this study, we describe the isolation and immortalisation of primary murine alveolar epithelial cells (mAEpC), as well as their epithelial differentiation and barrier properties when grown on Transwell® inserts. Like human alveolar epithelial cells (hAEpC), mAEpC transdifferentiate in vitro from an alveolar type II (ATII) phenotype to an ATI-like phenotype and exhibit features of the air–blood barrier, such as the establishment of a thin monolayer with functional tight junctions (TJs). This is demonstrated by the expression of TJ proteins (ZO-1 and occludin) and the development of high transepithelial electrical resistance (TEER), peaking at 1800ωcm 2 . Transport across the air–blood barrier, for general toxicity assessments or preclinical drug development, is typically studied in mice. The aim of this work was the generation of novel immortalised murine lung cell lines, to help meet Three Rs requirements in experimental testing and research. To achieve this goal, we lentivirally transduced mAEpC of two different mouse strains with a library of 33 proliferation-promoting genes. With this immortalisation approach, we obtained two murine alveolar epithelial lentivirus-immortalised (mAELVi) cell lines. Both showed similar TJ protein localisation, but exhibited less prominent barrier properties (TEERmax ~250Ωcm 2 ) when compared to their primary counterparts. While mAEpC demonstrated their suitability for use in the assessment of paracellular transport rates, mAELVi cells could potentially replace mice for the prediction of acute inhalation toxicity during early ADMET studies.