hESC/iPSC derived epithelial lineages. (A) Representative the epithelial differentiation process of hESCs or hiPSCs through supplement of RA and BMP4 in N2 medium for 7 days, then either changing into DSFM directly or passaging at a split ratio of 1:3 for culture continuously. Scale bar corresponds to 200 μm. (B) qPCR of representative experiment showing the down regulation of ESC-specific transcription factor (Oct4) and up regulation of keratinized epithelial markers (K18, p63, K19, CD29, K14). (C) Western blot for Oct4, p63, and K18 of lysates from H1 or ifhU1-iPSCs derived epithelial cells; GAPDH is the loading control. (D) Phase contrast captures and immunofluorescence staining [IF: p63 (red), K14 (green), DAPI (blue)] of epithelial sheets derived from H1 and ifhU1-iPSCs at D7 and D21. Scale bars correspond to 2000 and 200 μm, respectively. (E) Scanning electron microscope (SEM) and transmission electron microscopy (TEM) images of H1-ESCs and ifhU1-iPSCs derived epithelial sheets at D7 and D14. White arrowheads indicate desmosomes between epithelial cells of the sheet. Scale bars in SEM and TEM correspond to 20 and 0.5 μm respectively.