Extended Data Fig. 1: Generation of human blastoids from naive human PSCs.
From: Blastocyst-like structures generated from human pluripotent stem cells
a, Representative immunofluorescence co-staining images of SOX2, GATA3 and GATA6 showing differentiation of naive human PSCs in human TSM (left) and HDM (right). Scale bars, 100 μm. b, Representative immunofluorescence co-staining images of SOX2, GATA3 and GATA6 showing differentiation of naive human PSCs in TSM followed by HDM (top) and HDM followed by TSM (bottom). Scale bars, 100 μm. c, A representative phase-contrast image showing the presence of ELCs, HLCs and TLCs in a day-7 codifferentiation culture. Green, purple and red dashed lines encircle a typical colony of ELCs, TLCs and HLCs, respectively. Scale bar, 100 μm. d, Representative phase-contrast images showing cell aggregates formed in media with different TSM and 5i/L/A ratios. Red arrowheads indicate blastocyst-like structures. Scale bars, 100 μm. e, Schematic of human blastoid formation from naive human PSCs using the TH method. f, Representative phase-contrast images of human blastoids (TH method). Scale bars, 100 μm. g, Representative phase-contrast images showing human blastoids generated from 5i/L/A-cultured HFF and BJ human iPS cells and PXGL-cultured WIBR3 human ES cells (HT method). Scale bar, 100 μm. h, Human blastoid formation efficiencies from 5i/L/A-cultured HFF human iPS cells (HT method, n = 6 biological replicates; TH method, n = 3 biological replicates), 5i/L/A-cultured BJ human iPS cells (HT method, n = 3 biological replicates) and PXGL-cultured WIBR3 human ES cells (HT method, n = 3 biological replicates). Data are mean ± s.d. i, Lengths and widths of human blastocysts (n = 13 biological replicates) and human blastoids (TH method, n = 36 biological replicates; HT method, n = 110 biological replicates). j, Total number of cells per human blastocyst (n = 9 biological replicates) and human blastoid (HT method, n = 52 biological replicates; TH method, n = 16 biological replicates). Box plots show median (centre line), 25th and 75th percentiles (bottom and top of box, respectively), and minimum and maximum values (bottom and top whisker, respectively). k, Morphological grading of human blastocysts (n = 11 biological replicates) and human blastoids (HT method, n = 97 biological replicates; TH method, n = 21 biological replicates). TE, trophectoderm. Images in a–d, f, g are representative of at least three independent experiments.
