Extended Data Fig. 8: In vitro differentiation of stem cells derived from human blastoids.

a, Representative phase-contrast images showing blastoid TSCs, as well as EVT-like cells and SCT-like cells differentiated from blastoid TSCs. Scale bars, 100 μm. b, Representative immunofluorescence images of GATA3, CK7 and CDH1 (trophectoderm markers), SDC1 and CGB (SCT markers) and HLA-G (an EVT marker) in EVT-like cells and SCT-like cells differentiated from blastoid TSCs. Scale bars, 100 μm. c, RT–qPCR results showing the relative expression levels of EVT (FN1, HLA-G, ITGA5, ITGA1) and SCT (CGA, CGB (detects all 6 CGB genes), PSG1, CSH1, CYP19A1) markers in blastoid TSCs, as well as in EVT-like and SCT-like cells differentiated from blastoid TSCs (n = 3 biological replicates, mean ± s.d.). d, A representative phase-contrast image showing a day-9 differentiation culture of blastoid nEND cells. Scale bar, 100 μm. e, f, Representative immunofluorescence co-staining images of COL6A1, FOXA1 and GATA6 in a day-9 differentiation culture of blastoid nEND cells. White dashed lines indicate FOXA1⁻COL6A1⁺GATA6⁻ cells (e) and FOXA1⁺COLA6A1⁻GATA6⁻ cells (f). Scale bars, 100 μm. g, RT–qPCR results showing relative expression levels of VE/YE and EXMC markers in day-4 and day-9 differentiation cultures of blastoid nEND cells. (n = 3 biological replicates; mean ± s.d.). h, A representative phase-contrast image showing the colony morphology of primed human ES cells converted from blastoid naive ES cells. Scale bar, 100 μm. i, Representative immunofluorescence images showing that primed human ES cells converted from blastoid naive ES cells stained positive for SOX2 and CD24. Scale bars, 100 μm. j, Representative H&E staining images showing that the teratomas formed by primed human ES cells contain tissues from all three embryonic germ layers. Scale bar, 100 μm. Images in a, b, d–f, h–j are representative of at least three independent experiments.

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