Fig. 1

hPSC differentiation into a hematoendothelial population. A One hiPSC cell line (A29) and two hESC cell lines (SA01 and H1) were differentiated following embryoid body (EB) formation for 84 h supplied with growth factors inducing mesoderm and hematoendothelial specification. CD144⁺-sorted cells were differentiated into endothelial and hematopoietic lineage in the presence of specific growth factors. B–E EBs were analyzed at 48 h and 84 h by RTqPCR and compared to the hPSCs preceding EB formation (0 h) for the stem cell genes POU5F1 (B) and NANOG (C), the mesoderm gene TBXT (D) and the hematoendothelial gene KDR (E). F At 84 h, EBs were dissociated enzymatically and analyzed by flow cytometry for the expression of hematoendothelial (CD309, CD34 and CD143), endothelial (CD144 and CD31) and hematopoietic markers (CD43, CD41 and CD45), n = 30 for every hPSC cell line and marker. G Representative flow cytometry analysis of CD143, CD309, CD34 and CD31 within the CD144+ population from 84 h-EBs. H–I 84 h-EBs were sorted based on the expression of CD144 and analyzed for the expression of the hematoendothelial transcription factors ETV2 (h) and RUNX1 (I) by RTqPCR, n = 5 independent experiments for each cell line. *P value < 0.05; **P value < 0.005, ***P value < 0.0005. Data are represented as mean ± SEM