Fig. 2

Characterization of hESC-derived HEPs by analysis of gene expression pattern and structural features during the development of hematopoiesis after the differentiation. A Fluorescence microscopy of developing the EBs at day 7 after the differentiation (bright field (left panel) and the merge of immunofluorescence staining (right panel), the colors of green and blue represent the staining for CD31 and nuclei, scale bar, 40 μm). B CLSM images of the immunostaining for CD34 and CD31 in the EBs at day 7 after the differentiation. The colors of green, red and blue represent the staining for CD31, CD34 and nuclei. (Original magnification, 20×, scale bar 40 μm). C Representative flow cytometry results showed the population of CD34⁺CD144⁺ and CD34⁺CD31⁺ HEPs at different days during the hematopoietic specification after the hematopoietic differentiation from hESCs. D Morphology of HUVECs and HEPs at day 6 post-sorting and cultured in EGM2 medium (10× magnification, left panel), and CLSM images of the immunostaining for VE-Cadherin. The colors of green and blue represent the staining for VE-Cadherin and nuclei. (Right panel, original magnification, 40×, scale bar 50 μm). E Tube formation by HUVECs and HEPs on Matrigel-coated plate in EGM2 medium. (20,000 cells/well in 12-well culture plates, photographs taken 12 h after the culture; 10× magnification, left panel). CLSM images of the immunostaining for CD31. (Right panel, 40×, scale bar 50 μm). F Gene expression analysis for arterial genes (SOX17, DLL4 and NOTCH1) and venous gene (EPHB4) during the development of the hematopoiesis after the hematopoietic differentiation from hESCs. Data represent the mean ± SEM. *p < 0.05, **p < 0.01, and ns (no significance)