Extended Data Fig. 10: Hematopoietic cell composition in heX-embryoids.
From: Modelling post-implantation human development to yolk sac blood emergence
(a) Expression of Hemoglobin ε+ and Hemoglobin γ+ cells in day 15 heX-embryoids. At day 15, areas of Hemoglobin ε-high, Hemoglobin γ-low cells are observed, suggesting more primitive-like hematopoiesis. Scale bar = 500 μm. Dotted box outlines inset shown to the right. A representative image of 3 biological replicates. (b) Expression of Hemoglobin ε+ and Hemoglobin γ+ cells in day 21 heX-embryoid. At day 21, areas of Hemoglobin ε-high, Hemoglobin γ-high cells are observed, suggesting more definitive-like hematopoiesis. Scale bar = 500 μm. Dotted box outlines inset shown to the right. A representative image of 3 biological replicates. (c) Violin plot showing the distribution of the ratios of Hemoglobin ε to Hemoglobin γ at each day. n[D15] = 3,188 cells, n[D21] = 4088. Counts are from three biological replicates. Central dotted line indicates median, smaller dotted lines above and below indicate quartiles. (d) Subclustering of cluster 11, the putative endothelial population, from the day 21 scRNA-seq data. Hierarchical subclustering identifies two major subpopulations within this cluster, corresponding to hemogenic (11.1) and non-hemogenic (11.2) endothelial subtypes. Marker genes for each population are shown in violin plots to the right. (e) Expression of hemogenic endothelial markers within the day 21 hematopoietic populations. Markers co-express mainly in the area annotated as “blood progenitors” in the hypergeometric statistical comparison. (f) Scatterplots showing the co-expression of hemogenic endothelial markers within the day 21 hematopoietic populations. (g) Immunofluorescence images of day 21 of culture shows co-expression of RUNX1 and VE-cad. Scale bar = 50 μm. (h) Scatterplots showing the co-expression of respective lineage markers within the day 21 hematopoietic population.
