Figure 1
Endothelial differentiation of monkey IPSCs.
(A) Schematic illustration of the differentiation method. Cynomolgus monkey induced pluripotent stem cells (cIPSCs) were induced to mesoderm commitment (Priming phase) and further differentiated into endothelial cells (Induction phase). (B) Immunostaining at day 3 of differentiation indicates the formation of a mesodermal intermediate stage, as shown by the expression of Brachyury/T. Scale bar: 50 μm. (C) Flow cytometry analysis at day 6 of differentiation reveals that approximately 7.5% of cells had differentiated into endothelial cells (CD144+), while 82.5% of cells formed vascular smooth muscle cells (CD140b+). (D) Immunostaining of cIPSC-ECs at day 13 of differentiation reveals expression of EC specific markers VECAD, PECAM1, and vWF. Scale bars: 100 μm. (E,F) Global transcriptome analysis during the differentiation of cIPSCs to endothelial cells. (E) Principal component projections of Cynomolgus monkey transcriptomes colored by differentiation time. The variability of the data set along principal component 1 is 57% and along principal component 2 is 17%. (F) Heat map of marker gene panels for pluripotency, mesoderm (including Wnt target genes), and endothelial cells. Rows represent genes, and columns represent Cynomolgus monkey cells at different differentiation time points. Row Z-score transformation was performed on mean log2 values (n = 3 replicates) for each gene, with blue denoting lower and red denoting higher expression levels compared to the average expression level. Hierarchical clustering of genes and samples is based on complete linkage and Pearson correlation distance.
