Fig. 1: Differentiation and characterisation of hPSC-derived Müllerian duct-like cells.

a Diagram of stepwise differentiation of hPSCs into MDLCs with chemicals. MDLCs Müllerian duct-like cells. b Light microscopic observation of morphology in each stage. Yellow scale bars, 500 μm. c Expression level of KRT8, VIM, WNT7A and SIX2 in three cell lines at each stage. hUiPSCs human urine-derived induced pluripotent stem cells, hMiPSCs human monocyte-derived induced pluripotent stem cells. (VIM, VIMENTIN; KRT8, KERATIN 8) (data represent mean ± SEM, N = 3. *P < 0.05; **P < 0.01; ***P < 0.001; ns (no significance), P > 0.05 by one-way ANOVA and unpaired, two-tailed Student’s t test). d Immunofluorescence examination of multiple markers, PAX2, VIM and KRT8 in each stage. White scale bars, 20 μm. DAPI (blue). e Verification of epithelial/mesenchymal hybrid features of CHIR 2d-treated cells by immunofluorescence and flow cytometry. (pan-CK, pan-Cytokeratin) Green scale bars, 40 μm. f RNA-seq analysis of IM cells and CHIR 2d-treated cells, gene expression profiling by heatmap; representative upregulated genes are listed in this figure. (P value < 0.01). g Biological process of CHIR 2d-treated cells shown by GO. h Karyotype of hPSC-derived MDLCs.