Fig. 4: iPSC-derived airway epithelial cells grow in ALI culture and can be used to detect CFTR-dependent ion flow.
From: A multimodal iPSC platform for cystic fibrosis drug testing
A Schematic of the generation of iPSC-derived ALI cultures via FACS-purified NGFR + airway basal cells. B mRNA expression of canonical airway epithelial markers within iPSC-derived ALI cultures, compared to non-CF HBEC ALI cultures (biological replicates from independent differentiations: n = 2 for Phe508del #1, n = 3 for Phe508del #2, n = 4 for Phe508del #3). Each point represents an individual experiment. C Example of immunolabeling of iPSC-derived Phe508del ALI cultures with antibodies against markers of multiciliated (acetylated-α-tubulin), mucus secreting (MUC5AC), and airway basal cells (KRT5) with nuclei counterstained with Hoechst. D Uniform Manifold Approximation and Projections (UMAPs) of scRNA-seq of non-CF iPSC-derived airway ALI cultures depicting Louvain clustering and violin plots demonstrating the expression of CFTR in the annotated clusters. E Equivalent current measurements of Phe508del ALI cultures (n = 6 per treatment). Measurements are shown at baseline and after ENaC inhibition, forskolin treatment, CFTR potentiation with Genistein, and CFTR inhibition (arrows). F Quantification of peak delta forskolin and CFTR-inhibition effects of assay results shown in (E) (n = 6 experimental replicates from independent wells of a differentiation). P values were calculated using paired Student’s t test. Lines and error bars represent mean ± standard error. Scale bars represent 50 µm.
