Extended Data Fig. 19: Blastoid formation capacity of 4CL naïve PSC, e4CL-day 5 cells and sorted 8CLC.
From: Rolling back human pluripotent stem cells to an eight-cell embryo-like stage
a. Representative phase contrast images of sorted 8CLC-derived blastoids from day 1 to day 5. n = 5 biological replicates. Scale, 100 μm. b. Representative phase contrast images showing self-organized blastoids from sorted 8CLC, stepwise e4CL-day 5 cells and 4CL-day 12 naïve ESC. H9 ESC were used in all cases. D, day. Representative of five independent experiments. Scale, 50 μm. c. Representative immunofluorescence images of self-organized blastoids from 4CL-day 12 naïve ESC (left panel) and stepwise e4CL-day 5 cells (right panel). OCT4 and GATA2 were used as ICM and trophectoderm markers, respectively. Nuclei were counterstained with DAPI (blue). Scale, 20 μm. H9 ESC were used for these experiments. D, day. Representative of three independent experiments. d. UMAP visualization based on Fig. 4g highlighting the averaged expression scores of gene signatures and examples for the epiblast lineage in sorted 8CLC-derived blastoids and human E5–E7 blastocysts20. EPI, epiblast. All reference datasets used in this study are summarized in Supplementary Table 8. e. UMAP visualization based on Fig. 4g highlighting the averaged expression scores of gene signatures and examples for hypoblast lineage in sorted 8CLC-derived blastoids and human E5–E7 blastocysts. HYPO, hypoblast. f. UMAP visualization based on Fig. 4g highlighting the averaged expression scores of gene signatures and examples for the trophectoderm lineage in sorted 8CLC-derived blastoids and human E5–E7 blastocysts. TE, trophectoderm.
