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[Preprint]. 2023 Feb 28:2023.02.27.530299. [Version 1] doi: 10.1101/2023.02.27.530299

Figure 1: Analysis of epithelial structure and cell types produced by BCi-NS1.1- and primary-derived HAE cultures.

Figure 1:

a. Diagrammatical representation of experimental design. HAE cultures were generated from either primary cells or the hTERT-expressing precursor cell line BCi-NS1.1. Cells were differentiated on Transwells for three weeks at air-liquid interface to generate three replicates of BCi-NS1.1-HAE cultures (derived from one donor), and three HAE cultures from three separate primary donors. b. Hematoxylin & Eosin (H&E) and Periodic Acid-Schiff (PAS)-Alcian blue staining of formalin-fixed paraffin-embedded transections of HAE cultures from either BCi-NS1.1 or primary precursor cells. All cross-sectional images are oriented with the basolateral surface of the culture on the bottom of the image and the apical surface on the top of the image. Scale bars = 150μm c. Cell type-specific immunohistochemical staining of transections of BCi-NS1.1- or primary-derived HAE cultures. DAPI (nuclei) stained in blue. Cytokeratin 5 (KRT5, basal cells), villin-1 (ciliated cells), CC10 (encoded by SCGB1A1) (secretory cells) and MUC5B (secretory cells) stained in green. MUC5AC (secretory cells) stained in red. Scale bars = 150μm. d-g. Quantification using Imaris software of cells stained positive for each cell-type specific marker, KRT5 (c), villin-1 (d), CC10 (e), and MUC (f) (sum of MUC5B and MUC5AC positive cells), represented as a percentage of the total phalloidin (actin)-stained area of the HAE cultures. h. Quantification using Imaris software of cells stained positive for MUC5B, MUC5AC, or both, represented as a percentage of total MUC+ cells.