Extended Data Fig. 8: Mesp1^der PSC situated in the ventral region of the dorsal aorta have long-term growth and induce hematopoiesis.

(A) Flow cytometry using hematopoietic tissues from recipients 6 months after transplantation with re-aggregates of E11.5 endothelium or adult cardiac endothelium or E11.5 PSCs. (B) Flow cytometry of pooled co-aggregates (UBC-GFP adult heart endothelial cells + E11.5 Mesp1-DsRed⁺ PSC) at 96 h. (C) (i) Confocal image showing PDGFRA expression in the E11.5 dorsal aorta of a Mesp1-DsRed embryo. (ii) Flow cytometry of the ventral and dorsal halves of the E11.5 dorsal aorta in Mesp1-DsRed embryos (n = 5). (D) (i) Schematic outlining the process of harvesting endothelial cells from E11.5 AGM or adult heart and Mesp1^der PSCs from the dorsal or ventral halves of the E11.5 aorta for CFU-F assays and co-aggregate cultures. (ii) CFU-F activity of Mesp1^der PSCs from the whole AGM or dorsal or ventral halves of the E11.5 aorta. (iii) Long-term replating of Mesp1^der PSCs from the whole AGM or dorsal or ventral halves of the E11.5 aorta. (iv) CFU-C potential of embryonic or adult endothelium co-aggregated with E11.5 Mesp1^der PSCs from either the dorsal or ventral halves of the aorta. (v) Percentage of GFP⁺ cells in peripheral blood of irradiated recipients 6 months after transplantation of co-aggregates (one co-aggregate for each adult irradiated recipient). CFU-C: colony-forming unit–culture; BFU-E: burst-forming unit–erythroid; CFU-GM: colony-forming unit–granulocyte/macrophage; CFU-GEMM: colony-forming unit–granulocyte/erythrocyte/macrophage/megakaryocyte. * p < 0.05, ** p < 0.01, *** p < 0.005; random effects Poisson regression was used to compare colony counts (D(ii), (iv)), ANOVA was used to compare donor chimerism (D(v)) and a linear mixed model was used to compare the growth curves (D(iii)). Data were derived from biologically independent samples, animals and experiments. Data represent mean ± S.D. The precise p values are listed in the source data.

Source data