Isolation, omics characterization and organotypic culture of alveolar type II pulmonary epithelial cells
Li, H.; Schuette, M.; Bober, M.; Kroll, T.; Frappart, L.; Bou About, G.; Lin, Y.-C.; Sorg, T.; Herault, Y.; Wierling, C.; Rinner, O.; Lange, B. M.; Ploubidou, A.
Show abstract
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.
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