Premium
A primary in vitro co‐culture model of the human alveolo‐capillary barrier to study lung injury and repair
Author(s) -
Hermanns Maria Iris,
Fuchs Sabine,
Wenzel Katja,
Bittinger Fernando,
Kirkpatrick C. James
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a210-b
Subject(s) - adherens junction , tight junction , alveolar epithelium , transdifferentiation , microbiology and biotechnology , cell culture , occludin , monolayer , chemistry , biology , cadherin , epithelium , pathology , cell , medicine , stem cell , biochemistry , genetics
Due to an increasing interest in pulmonary toxicology and pathobiology, there is a need to develop cell culture models of the alveolar‐capillary barrier. We established a co‐culture system of human pulmonary microvascular endothelial cells (HPMEC) and human alveolar epithelial cells, consisting of type II (hATII) and type I‐like (hATI‐like) cells, on opposite sides of a permeable filter support. Primary isolated hATII cells were seeded on the top surface of collagen typ I‐ coated Transwell filter units (24‐well filter plate) and grown to confluence simultaneously with HPMEC on the lower surface over ten to fourteen days. Within 10 days of co‐culture the hATII cells partly transdifferentiated to ATI‐like cells, demonstrated through morphological changes from a cuboidal to a flattened morphology of the cells, the loss of cell type specific organells (lamellar bodies) and an increased mRNA expression of RAGE and AQP‐5. hATII and hATI‐like cells established a contact inhibited monolayer, showing a circumferential staining of the tight junction and the adherens junction proteins (ZO‐1, Occludin, E‐Cadherin, alpha‐ and β‐Catenin). The generation of a polarized epithelial cell monolayer resulted in trans‐monolayer resistance values of 1250±350 Ohm*cm 2 and trans‐bilayer resistance values of 1730±460 Ohm*cm 2 from day 7 to day 10. HPMEC on the opposite side of the permeable 24‐well filter plate developed monolayer with tight and adherens junctions. Our primary co‐culture model will be suitable to examine interactions of both alveolar epithelium and microvascular endothelium in the pathogenesis of lung injury and the following repair processes.