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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Nat Med. 2015 Oct 26;21(11):1364–1371. doi: 10.1038/nm.3973

Figure 1. Induction of polarized organoids from human pluripotent stem cells.

Figure 1

(a) A schematic diagram of the protocol for growing pancreatic lineage committed pluripotent stem cells (PSCs) on a 3D platform. PTOM refers to Pancreatic Progenitor and Tumor organoid Media and POMM refers to Pancreatic Organoid Maintenance Media. See online methods for details. (b) Phase morphology of day 16 organoids (left panel) with insert representing higher magnification image of one organoid. H&E staining of one organoid (right panel). (c) Confocal images of day 16 organoids immunostained for basal markers COLLAGEN IV (COLIV) or LAMININ α5 (LAMA5) (red), tight junction marker (ZO1, green), cell-cell junction marker, E-CADHERIN (ECAD, green) and DAPI (blue). (d) Transmission electron micrograph of cells from day 16 3D organoids. Upper panel: apical region of epithelia, arrowhead pointing to an electron dense region representing tight junctions. Lower panel: basal region of polarized epithelia with the arrowhead pointing to basement membrane. Scale bars, 0.5 µm. (e) Staining for Ki67 at different days in 3D culture. DAPI, blue; Ki67, red. (f) Cell polarization and apoptosis during pancreatic organoid morphogenesis. Top panel: apoptosis marker Cleaved Capase-3 (CC3, red), and apical maker MUCIN1 (MUC1, green), DAPI (blue). Bottom panel: basal and apical markers COLIV (red) and ZO1 (green), respectively in progenitor organoids from day 2 to day 10 in 3D culture. (g) Maintenance of polarity upon serial passaging of organoids as shown by staining for ACTIN (green), COL IV (red) and ECAD (purple). Scale bars represent 50 µm, unless specified otherwise.