Fig. 3.
Development of Runx1hi CD41+ CD45+ HSPCs is promoted by HOXB4. A Dissociated day 6 EBs were co-cultivated on OP9 stroma for 5 days (EBd6/OP9d5) and analyzed for surface expression of CD41 and CD45. B Flow cytometry-based separationof EBd6/OP9d5 cells in Venus(Runx1) negative (neg), intermediate (int) and high (hi) expressing subpopulations. A representative result of 5 independent ESC differentiations is shown. The mean values of the percentages of Venus(Runx1)+ cells ± SD were: 16.6 ± 8.4% (HOXconst) versus 20 ± 16% (control) for all Runx1+ cells. Within the Venus(Runx1)+ population, Runx1hi cells accounted for 14.8 ± 8.8%. No Runx1hi were detected without ectopic HOXB4 expression. C Venus(Runx1)+ hematopoietic cell clusters arising during OP9 co-culture. Phase contrast (PC, upper panel) and fluorescence (lower panel) images are shown.