Fig. 3: Induction of blastocyst-like structures from chemically induced totipotent-like cells.
a, Bright-field image of blastoids at S4. Scale bar, 100 μm. Similar images were obtained in at least three independent experiments. b,c, Immunofluorescent analysis showing the expression of OCT4 and CDX2 in blastoids at S4 (b) and E4.5 blastocysts (c). n = 9 S4 blastoids from 6 experiments, n = 3 E4.5 blastocysts from 2 experiments. Scale bars, 20 μm. d,e, Immunofluorescent analysis showing the expression of OCT4 and SOX17 in blastoids at S4 (d) and E4.5 blastocysts (e). n = 7 S4 blastoids from 5 experiments, n = 3 E4.5 blastocysts from 2 experiments. Scale bars, 20 μm. f, Histograms showing the distribution of the diameters of S4 blastoids and E4.5 blastocysts. n = 37 S4 blastoids, n = 27 E4.5 blastocysts. The vertical dashed line denotes the mean of the group. g, Histograms showing the total cell number of S4 blastoids and E4.5 blastocysts. Data are presented as mean ± s.d. n = 34 S4 blastoids, n = 26 E4.5 blastocysts. n.s., non-significant (two-sided Student’s t-test). h, Histograms showing the indicated cell number of S4 blastoids and E4.5 blastocysts. Data are presented as mean ± s.d. n = 62 S4 blastoids, n = 47 E4.5 blastocysts. *P < 0.05; n.s., non-significant (two-sided Student’s t-test). i, Dot plots showing the average levels and proportion of cells expressing lineage-specific marker genes in blastoids at S4. Lineage-specific marker genes for EPI, TE and PrE in E4.5 mouse blastocysts were identified using published single-cell transcriptomic datasets (GSE123046). Expressions of these marker genes in EPI, TE and PrE from E4.5 mouse blastocysts are shown as the control. j, UMAP analysis of transcriptome of cells from blastoids at S4. These samples were integrated with published single-cell transcriptomic datasets from E4.5 mouse blastocysts (GSE123046).
