Fig. 6

Characterization of spatial self-organization within the hiPSC colony with or without the ring-barrier using the ring culture system under the influence of endodermal differentiation conditions. A Experimental design for endodermal differentiation based on the ring culture system. B Representative morphologies are shown at different time points during endodermal differentiation within the hiPSC colony with or without the ring-barrier. Scale bar, 500 μm. C Representative images in the XY plane showing expression of pluripotency (OCT3/4) and endodermal (FOXA2, SOX17) markers. Images are shown as one quarter of a colony. Panels a1–r1 are the tomograms sectioned at the XZ plane (yellow dashed lines) in panels a-r. White dashed lines indicate location of the ring culture system. Nuclei were stained with DAPI. Scale bar, 50 μm. All experiments were repeated independently at least three times with similar results