ena-STUDY-Alacris Theranostics GmbH-18-12-2019-14:06:31:071-1310
Isolation, 'omics characterization and organotypic culture of alveolar type II pulmonary epithelial cells
Alveolar type II (AT2) epithelial cells include the stem cells of the pulmonary alveoli, functioning in lung homeostasis and post-injury repair, but also mediate biophysical and immunological aspects of lung physiology. AT2 cells have been characterized primarily in situ, in transgenic mouse models. Here, we report new methodology for their isolation, “omics” characterization and stroma-cell-free organotypic culture. 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 proteins. 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 alveolar cells and of lung cancer mouse models.
Alveolar type II (AT2) epithelial cells include the stem cells of the pulmonary alveoli, functioning in lung homeostasis and post-injury repair, but also mediate biophysical and immunological aspects of lung physiology. AT2 cells have been characterized primarily in situ, in transgenic mouse models. Here, we report new methodology for their isolation, “omics” characterization and stroma-cell-free organotypic culture. 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 proteins. 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 alveolar cells and of lung cancer mouse models.