Figure 1.

Differentiation of patient-derived iPSCs to lung progenitor cells

(A) Schematic of differentiation protocol and timeline. Human iPSCs were directed to definitive endoderm and passaged onto 96 well plates during the anterior foregut endoderm differentiation. The submerged cultures were differentiated for 10 more days into lung progenitor cells (stage 3b; Wong et al., 2015).

(B) Immunofluorescence images of submerged cultures. Scale bar, 80 μm. Most cells stained positive for TTF1 (NKX2-1) and SOX9. Negative controls are shown in Figure S1.

(C) Principal-component analysis (PCA) comparing iPSC lines, submerged lung progenitor cultures differentiated from iPSC lines, and primary bronchial cultures. Both CF and non-CF (including mutation-corrected) iPSC lines were studied.

(D) Heatmap of gene expression clustered according to cell type using marker genes (Deprez et al., 2020). The columns correspond to different donors and whether lines are CF or non-CF (including mutation corrected [MC]). Columns are also clustered as an embryonic stem cell line (H1), iPSC lines, lung progenitors differentiated from iPSC lines as submerged cultures (SC), and primary human bronchial epithelial (HBE) cultures. Relative CFTR expression across cultures is shown in the bottom row of the heatmap.