Fig. 1

The number and functions of BM EPCs from patients with MDS. HD and patients with de novo AML were enrolled as controls. A, left The EPC phenotype was characterized by demonstrating positive expression of CD34, CD309 and CD133 by flow cytometry. The A, right percentage of BM EPCs in precultivated BMMNCs was analysed. B Representative images (scale bars represent 200 µm) of typical cultured BM EPCs collected at day 7 of culture from HDs and L-MDS, H-MDS and AML patients were characterized by double-positive staining (merged in yellow) with DiI-AcLDL (red) and FITC-UEA I (green) (original magnification, 10×). C Quantification of double-positive EPCs/field of view (merged in yellow) stained with DiI-AcLDL (red) and FITC-UEA I (green) at day 7 of culture (original magnification, 10×). D Representative images (scale bars represent 200 µm) of tube formation (pixels of tubes per field of view) by BM EPCs at day 7 of culture (original magnification, 10×). E Quantification of tube length (pixels of tubes per field of view) of BM EPCs at day 7 of culture (original magnification, 10×). F BM EPCs at day 7 of culture were cultured in a transwell chamber for 24 h, fixed and then stained with crystal violet. Representative images (scale bars represent 200 µm) of migrated cells on the bottom surface of the membrane (original magnification, 10×). G The number of migrated BM EPCs per field of view was compared (original magnification, 10×). Three power fields were randomly counted and averaged per sample. Statistical analyses were performed using the Mann–Whitney U test. Data are presented as the means ± SEM (*P ≤ 0.05, **P ≤ 0.005, ***P ≤ 0.001). AML Acute myeloid leukaemia, BM Bone marrow, BMMNCs Bone marrow mononuclear cells, EPCs Endothelial progenitor cells, DiI-Ac-LDL DiI-acetylated low-density lipoprotein, FITC-UEA I FITC-labelled Ulex Europaeus Agglutinin I, HD Healthy donor, H-MDS Higher-risk myelodysplastic syndromes, L-MDS Lower-risk myelodysplastic syndromes