Additional file 5: Figure S2. of Optimizing mesoderm progenitor selection and three-dimensional microniche culture allows highly efficient endothelial differentiation and ischemic tissue repair from human pluripotent stem cells

3D microniche dramatically improved endothelial differentiation efficiency from hiPSCs. (a and b) Immunostaining showing CD31 expression in 2D (a) and 3D GM (b) on differentiated day 12 respectively. Scale bars: 100 μm. (c) Percentage of CD31+ cells on differentiated day 12 in 2D and 3D GM (n = 3, *** p < 0.001). Figure S3. Signaling pathway analysis of enriched genes in 3D conditions. (a) Heatmaps of 3D enriched genes belonging to the following GO classes: extracellular matrix, vasculature development, regeneration, response to steroid hormone, response to wounding, and regulation to cell proliferation. (b) Q-PCR validation of representative endothelial marker genes enriched in 3D GM differentiation in iPSCs (n = 3, ** p < 0.005, *** p < 0.001, 3D versus 2D; t test), related to Fig. 5. Figure S4. In vivo tumorigenicity test of MESP1+ cells. (a) Representative images showing teratoma formation in the right leg of the mouse injected with undifferentiated MESP1-mTomato cells (red circle). No teratoma was found in immune-deficient mice injected with the same number of differentiated MESP1-mTomato+ cells. (b) Table summarizing the teratoma formation result from undifferentiated MESP1-mTomato and differentiated MESP1-mTomato+ cells. Similar results were obtained with human iPSCs and other ESC lines (H1 and H7) (data not shown). (ZIP 10253 kb)