Fig. 4

Characterization of spatial self-organization within the hiPSC colony with or without the ring-barrier in the absence of exogenous differentiation factors. Immunofluorescent images of a pluripotency markers (OCT3/4 and SOX2) and differentiation markers including ectoderm (PAX6), mesoderm (BRACHYURY), endoderm (SOX17), and extra-embryonic trophoblast (CDX2). The hiPSC colonies were compared after further exposure to HA in culture conditions without the ring-barrier. Red dashed lines indicate location of the ring culture system. Scale bar, 500 μm. All experiments were repeated independently at least three times with similar results