Fig. 1: ETiX embryoids recapitulate developmental milestones of the natural mouse embryo up to E8.5.
From: Embryo model completes gastrulation to neurulation and organogenesis
a, Schematic of ETiX embryoid formation. ETiX embryoids are formed by aggregating ES cells, TS cells and ES cells transiently expressing GATA4, and by day 4 they generate structures that resemble post-implantation stage natural E5.5 embryos. They subsequently develop to gastrulation (E6.5/ETiX day 5 (ETiX5)) and neurulation (E8.0/ETiX day 7 (ETiX7)) stages before initiating organogenesis (E8.5/ETiX day 8 (ETiX8)). b,c, Bright-field images of natural mouse embryos (b) and ETiX embryoids (c) at different timepoints highlighting morphological similarities (n = 1,197 ETiX4, 237 ETiX5, 170 ETiX6, 100 ETiX7 and 40 ETiX8, from 17 independent experiments). Scale bars, 100 μm. d, Uniform manifold approximation and projection (UMAP) analysis of scRNA-seq data at indicated timepoints for natural embryos at E6.5, E7.5 and E8.5 and ETiX embryoids at days 5, 6 and 8 (n = 29 ETiX5, 10 ETiX6, 7 ETiX8, 12 E6.5, 14 E7.5 and 9 E8.5) analysed by inDrops sequencing. e, Single-cell inDrops RNA-seq UMAP annotated to show cell types identified in natural embryos and ETiX embryoids. AVE, anterior visceral endoderm; CLE, caudal lateral epiblast; NMP, neuromesodermal progenitors; PLPM, posterior lateral plate mesoderm. f, Annotated and combined UMAP of natural embryos cultured ex utero and collected at indicated timepoints (E7.5, E8, E8.5, E8.75 and E9.5) and ETiX embryoids (day 6 and day 8) individually labelled and analysed by tiny-sci-RNA-seq. n = 8 natural embryos ranging from E7.5 to E9.5, n = 3 ETiX6, 2 failed ETiX6, 5 ETiX8 and 4 failed ETiX8.
