Fig. 2

Differentiation of hESC-derived NCCs into corneal endothelial cells. a Schematic illustration of the differentiation conditions used to generate corneal endothelial cells from NCCs. b The hexagonal morphology of iCECs differentiated from hESC-derived NCCs. Scale bar: 50 μm. c Immunofluorescence staining showing that iCECs expressed ZO-1, Na⁺ K⁺ ATPase, AQP1 and N-cadherin. Nuclei were stained with DAPI. Scale bars: 50 μm. d Quantification of the percentage of ZO-1+, Na⁺ K⁺ ATPase+, AQP1 + and N-cadherin + cells at day 24 of differentiation. e Flow cytometry analysis of ZO-1/Na⁺ K⁺ ATPase, N-cadherin and CD166 expression in iCECs. n = 3