Isolation, ‘omics characterization and organotypic culture of alveolar type II pulmonary epithelial cells

Li, Huaibiao; Schütte, Moritz; Bober, Magdalena; Kroll, Torsten; Frappart, Lucien; About, Ghina Bou; Lin, Yu-Chieh; Sorg, Tania; Herault, Yann; Wierling, Christoph; Rinner, Oliver; Lange, Bodo MH; Ploubidou, Aspasia

The alveolar type II (AT2) epithelial cell fraction includes the stem cells of the pulmonary alveoli, functioning in lung homeostasis and post-injury repair. AT2 cells have been characterized primarily in situ, in transgenic mouse models. We report a new methodology for their isolation, their “omics” characterization and stroma-cell-free organotypic culture. Our multi-omics analysis identified high expression of genes involved in oxidative phosphorylation and of AP-1 components, as well as new phosphorylation sites in AT2 biomarkers. Furthermore, we show that supplementation with KGF, FGF10 & HGF suffices for the in vitro proliferation of AT2 cells and formation of alveolar organoids, suggesting that AT2-based organotypic development depends on ligands of the c-Met and FGFR2 receptors. The reported methodology and in-depth molecular characterization provide new tools for the in vitro and in vivo functional analysis of pulmonary cells and of mouse models of lung disease.