Fig. 1

Direct differentiation of hESCs into neural crest cells. a hESC-derived NCCs have the typical morphology of neural crest cells. Scale bars: 100 μm. b Immunofluorescence staining showing that hESC-derived NCCs expressed P75, NESTIN, AP2α, and SOX10. Nuclei were stained with DAPI. Scale bar: 50 μm. c Quantification of the percentage of P75+, NESTIN+, AP2α + and SOX10 + cells at day 12 of differentiation. d Flow cytometry analysis of P75, HNK1, NESTIN and SOX2 expression in hESC-derived NCCs. These images are representative of n = 3 biological replicates. e The expression of pluripotency genes (POU5F1, NANOG) and NCCs markers (B3GATI, NESTIN, NGFR, SOX10, TFAP2A) in hESCs and hESC-derived NCCs was detected by qPCR. n = 3, ^*p < 0.05, ^**p < 0.01, ^***p < 0.001, ^****p < 0.0001. f Flow cytometry analysis of CD90, CD105, CD73, CD29, CD19 and CD34 expression in hESC-NCC-derived mesenchymal stem cells. n = 3. g hESC-NCCs could be differentiated into mesenchymal derivatives, including Oil Red O-stained adipocytes (red), Alizarin red-stained osteocytes (red), and Alcian blue-stained chondrocytes (blue). Scale bars, 100 μm. h Immunofluorescence staining showing that hESC-derived NCCs could be differentiated into peripherin neurons, which expressed TUJ1 and Peripherin. Nuclei were stained with DAPI. Scale bar: 50 μm. i S100β + Schwann cells were detected in differentiated hESC-derived NCCs. Nuclei were stained with DAPI. Scale bar: 50 μm