Fig. 5: Haematopoietic programme characterization in heX-embryoids.
From: Modelling post-implantation human development to yolk sac blood emergence
a, IF of day 12 erythroid-like progenitors. b, IF of day 12 myeloid-like progenitors. c, IF of day 12 megakaryocyte-like progenitors. d, IF of day 17 multilineage haematopoietic foci. e, IF of day 17 uni-lineage haematopoietic foci. f, Results of a CFU methylcellulose assay seeded from day 21 cultures. Counts from n = 3 biological replicates shown. Example colonies are shown. CFU-E, colony forming unit-erythroid; BFU-E, burst forming unit-erythroid; GM, colony forming unit-granulocyte, macrophage; GEMM, colony forming unit-granulocyte, erythrocyte, macrophage, megakaryocyte. g, UMAP of day 21 populations. Boxed area shows populations with similarity to in vivo haematopoietic lineages. Hypergeometric statistical comparison to ref. 23 for these clusters is shown. For hypergeometric tests, we performed the Benjamini–Hochberg correction for multiple comparisons in all comparisons. Tests were one-sided. h, IF of day 12 and 21 cultures, showing representative erythroid-like colonies with primitive (high Hb ε, low Hb γ) characteristics on day 12 and definitive (high Hb γ, high Hb ε) characteristics on day 21. i, qPCR showing change HBE and HBG expression ratio across the indicated days. **P = 0.0012 (confidence interval 95%). P was calculated using one-way ANOVA with Tukey’s multiple comparisons test between days 12 and 21. n = 3 biological replicates sampled per day. j, Flow cytometry scatterplots showing erythroid- (CD235ab+), myeloid- (CD33+) and megakaryocyte-like (CD42b+) populations within the CD43+ population of day 21 cultures. Bar plots show cells identified within the CD43+ population in eight biological replicates. k, Flow cytometry analysis of week 3 culture for neutrophil-like cells, pregated for CD45+ cells. The bar plot shows the percentage of CD31+CD15+ cells in three biological replicates. l, Flow cytometry analysis of week 2 culture for lymphoid-like progeny, pregated for CD45+CD117+ cells. The bar plot shows the percentage of CD117+CD43+CD7+ cells in three biological replicates. m, Flow cytometry analysis of week 3 culture for natural killer-like cells, pregated for CD45+VLA-4+ cells. The bar plot shows the percentage of CD56+VLA-4+ cells in four biological replicates. Error bars are ±s.e.m. Scale bars, 100 µm.
