Fig. 5

Marker expression and functional proliferative and regenerative properties of the ABCG2-positive hPSC-LSCs cultured in the novel maintenance condition. a Morphology and p63α/ABCG2 expression pattern of hPSC-LSCs cultured in CnT-07+ENRC up to passages 0, 1, and 10. Cell nuclei counterstained with DAPI (blue). Scale bars, black = 200 μm, white = 100 μm. b Population doublings and c population doubling times of freshly differentiated hPSC-LSCs up to passage 5 and cryopreserved hPSC-LSCs up to passage 10. d Characterization of expression of the quiescence marker p27 in relation to ABCG2 and ∆Np63 at d11, as well as after continued culture in CnT-07+ENRC or CnT-30 at d24. Scale bars, 100 μm. e Hematoxylin and eosin-stained sections of CnT-30-cultured Regea08/017 and CnT-07+ENRC-cultured Regea11/013 hPSC-LSCs after 1-week and 2-week time points in the ex vivo porcine cornea model, respectively. Asterisk points out migrating cells. Scale bars, 500 μm and 100 μm, for all images in the panel. Data are presented for the hESC lines Regea11/013 (a–c, e) and Regea08/017 (d, e)