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Additional file 6. Supplementary figure 6. Representative flow cytometry histograms and dot plots of hPSC-BCs from A29 (top), SA01 (center) and H1 (below) cell lines for the expression of hematoendothelial (CD309, CD143 and CD34), endothelial (CD144 and CD31) and hematopoietic markers (CD43, CD45 and CD41) as (A) simple and (B) double labeling.

Additional file 7. Supplementary figure 7. Relative mRNA expression of the hemoglobin subunit epsilon-1 (HBE1), gamma-1 (HBG1) and beta-1 (HBB1) in the BFU-E and CFU-GEMM derived from every hPSC-cell line compared to mRNAs from human embryonic liver.

Additional file 17. Supplementary table 8. Average percentage of positive cells for the endothelial and hematopoietic markers analyzed by flow cytometry in hPSC-BCs. Supplementary to Figure 4B-C.

Additional file 5. Supplementary figure 5. (A) Representative macroscopic images (left panels) of a mouse dorsal chamber without (superior panel) and with ischemia (inferior panel) at day 5 and day 29 post-injection of hPSC-ECs. Right panels show the corresponding treated images used for quantification of the area occupied by blood vessels. (B) Representative confocal image at day 19 post-injection of a mouse dorsal skinfold chamber with ischemia, injected with mcherry-hPSC-ECs (red, white arrows). Vessel perfusion was observed thanks to the injection of dextran 70 kDa-FITC (green). Scale bar: 200 μm.

Additional file 13. Supplementary table 4. Oligonucleotides used in qPCR for the detection of human DNA.

Additional file 3. Supplementary figure 3. Arterial and vein differentiation of hPSC-ECs. (A) Differentiation scheme of hPSC-EB-CD144+ into hPSC-ECs in the presence or absence of DAPT and representative phase-contrast microscopy images after differentiation. Scale Bar 500 μm. Flow cytometry analysis of the endothelial phenotype of hPSC-ECs in the presence and absence of DAPT. (B) Confocal immunofluorescent image of Matrigel networks formed by hPSC-ECs. CD31 in green and DAPI in blue. Scale Bar: 100 μm.

Additional file 10. Supplementary table 1. List of antibodies used in flow cytometry and immunofluorescence analysis.

Additional file 16. Supplementary table 7. Summary of average ± SD values of in vitro endothelial characterization tests: Matrigel tube formation assay, TNFα activation assay, eNOS expression and NO production. Supplementary to Figure 3A-D.

Additional file 19. Supplementary table 10. Contains list of significantly hematopoietic and endothelial Gene Ontology Biological Process enrichment in hPSC-EB-CD144+, hPSC-BCs, hPSC-BC-CD144+CD45+ and hPSC-ECs compare to EL cell populations. Enrichment analysis was performed using Gene Set Enrichment Analysis software v4.1.0 and probing the c5.go.bp.v7.4 collection of the Molecular Signatures Database (MSigDB). Number of genes involved in the gene set, Enrichment score (ES), normalized enrichment score (NES) and nominal P-value are given in the supplementary Table 6. All selected Gene Ontology Biological Process enrichment show a P value ≤ 0.05.

Additional file 14. Supplementary table 5. Average percentage ± SD of positive cells for the endothelial and hematopoietic markers analyzed by flow cytometry in hPSC-EBs. Supplementary to Figure 1F.

Additional file 8. Supplementary figure 8. Counting of differential transcripts between undifferentiated hESCs, differentiated hESCs and EL cell populations. P

Additional file 1. Supplementary figure 1. (A) Representative image of the flow cytometry analysis of 84h- hPSC-EBs for hematoendothelial (CD309, CD143 and CD34), endothelial (CD144 and CD31) and hematopoietic markers (CD43, CD45 and CD41) for every hPSC line. (B) Representative analysis of hematoendothelial, endothelial and hematopoietic markers within the positive population for CD144, CD143, CD309, CD34 and CD31 for A29-EBs.

Additional file 4. Supplementary figure 4. (A) Representative histograms of the flow cytometry analysis of eNOS expression in hPSC-ECs and ECFC (B) Representative histograms of the flow cytometry analysis of the probe DAF-FM for NO detection in hPSC-EC and ECFC in the presence and absence of LPS and SNAP.

Additional file 15. Supplementary table 6. Average percentage ± SD of positive cells for the endothelial and hematopoietic markers analyzed by flow cytometry in hPSC-ECs during consecutive passages. Supplementary to Figure 2D.

Additional file 18. Supplementary table 9. Contains a list of significantly hematopoietic and endothelial Gene Ontology Biological Process enrichment in hPSC-EB-CD144+, hPSC-BCs, hPSC-BC-CD144+CD45+ and hPSC-ECs vs undifferentiated hESC. Enrichment analysis was performed using Gene Set Enrichment Analysis software v4.1.0 and probing the c5.go.bp.v7.4 collection of the Molecular Signatures Database (MSigDB ). Number of genes involved in the gene set, Enrichment score (ES), normalized enrichment score (NES) and nominal P-value are given in the supplementary Table. All selected Gene Ontology Biological Process enrichment show a P value ≤ 0.05.

Additional file 9. Supplementary figure 9. (A) Heatmap depicting mean level expression of POU5F1, NANOG, KDR, CDH5, CD34, SPN, ITGA2B and PTPRC genes in undifferentiated hPSC (H1 cell line), in hPSC-EB-CD144+, hPSC-BCs and hPSC-BC-CD144+CD45+ and hPSC-ECs derived populations obtained by gene microarray analysis (GeneChip® HumanGene 2.0 ST; Affymetrix). Mean level expression of selected genes are also visualizing in CD144+CD45-CD34+ EL-EC, CD144+CD45lowCD34+ EL-pre-HSC, CD144-CD45lowCD34+ EL-HSC/HP and CD144-CD45highCD34-EL-mature HC EL cell subpopulations for comparison. Heatmap revealed the loss of pluripotency during hPSC-EB-CD144+ formation and subsequent hPSC- BCs and hPSC-ECs derivation and distinct endothelial and hematopoietic signatures in hPSC-EB-CD144+, hPSCBCs, hPSC-BC-CD144+CD45+ and hPSC-ECs derived populations. (B) Heatmap depicting mean level expression of embryonic HBE1 (Hemoglobin ε), fetal HBG1 (Hemoglobin γ), and adult HBB (hemoglobin β) genes in undifferentiated hPSC (H1 cell line), in hPSC-EB-CD144+, hPSC-BCs and hPSC-BC-CD144+CD45+ and hPSC-ECs derived populations obtained by gene microarray analysis (GeneChip® HumanGene 2.0 ST; Affymetrix). Mean level expression of selected genes are also visualizing in CD144+CD45lowCD34+ EL-pre-HSC, CD144-CD45lowCD34+ EL-HSCs/HPs and CD144-CD45highCD34- EL-mature HCs EL cell subpopulations for comparison. Heatmap revealed that hPSC-BCs, hPSC-BC-CD144+CD45+ contained principally embryonic erythrocytes and to lesser extent definitive erythrocytes. Undifferentiated hESC (H1) , hESC-derived cell populations (n=3) and EL cell populations (n=4) After normalization, for each cell population, a Tukey's Biweight Robust Mean is calculated from log2 expression values. For each gene analyzed the most highly expressed transcript, is in red, the lowest level expressed transcript is in green. ID, Identifiant Affymetrix; Undiff, undifferentiated.

Additional file 2. Supplementary figure 2. (A) Representative phase-contrast images of hPSC-derived endothelial cells (hPSC-ECs) from passage (p) 1 to 5 derived from H1-CD144+-EBs. Scale bar 500 μm. (B) Representative flow cytometry histograms of hPSC-ECs from A29 (top), SA01 (center) and H1 (below) cell lines at p1 and p3 for the expression of hematoendothelial (CD309, CD143 and CD34), endothelial (CD144 and CD31) and hematopoietic markers (CD43, CD45 and CD41).