Extended Data Fig. 5: Monkey embryoids induction in vitro from monkey ESC-2# and iPSCs.
From: Modelling late gastrulation in stem cell-derived monkey embryo models
a, Representative brightfield images of dynamic induction process of monkey blastoid from D0 to D7 and IVC1-treated monkey embryoids on D9 in aggrewell. Scale bars, 100 μm. b, Representative brightfield images of lateral viewed monkey embryoids from D13 to D25. A, anterior; P, posterior. Scale bars, 200 μm. c, Developmental efficiency of 3.5 cm dish-cultured monkey embryoids from D13 to D17 (n = 3 independent experiments for each group). Monkey embryoids with clear embryonic disc-like structure including epiblast, amnion cavity and yolk sac cavity were calculated for statistical analysis. Each dot represents one repeat. d, Development efficiency of single cultured monkey embryoids from D17 to D25. Monkey embryoids with clear embryonic disc-like structure were calculated for statistical analysis (53 monkey embryoids on D17 from 3 independent experiments were used to track developmental efficiency to D25). e, Measurement of long diameters of whole embryo and embryonic disc length of monkey embryoids from D13 to D25 respectively (n = 14 for D13 and D15 groups; n = 9 for D17 group; n = 13 for D20 group; n = 11 for D23 group; n = 12 for D25 group from 4 independent experiments). f, Representative 3D images (upper) and monolayer (lower) showing the typical lateral view of monkey embryoids on D17 by light sheet microscope analysis after whole-mount IF staining with OCT4 (gray), TFAP2A (red), and SOX17 (green) (Supplementary Video 12). OCT4, a marker for epiblast; TFAP2A, a marker for amnion cells; SOX17, a marker for yolk sac. Scale bars, 200 μm. g, Representative 3D images showing the typical lateral view and dorsal view of monkey embryoids on D20 by light sheet microscope analysis after whole-mount IF staining with TBXT (gray) and DAPI (Supplementary Video 13). TBXT, a marker for PS (primitive streak). Scale bars, 200 μm. h, Representative brightfield images of dynamic induction process of monkey blastoid from D0 to D7 and IVC1-treated monkey embryoids on D9 in aggrewell. Scale bars, 100 μm. i, Representative brightfield images of lateral viewed monkey embryoids from D13 to D25. A, anterior; P, posterior. Scale bars, 200 μm. j, Developmental efficiency of 3.5 cm dish-cultured monkey embryoids from D13 to D17 (n = 5 independent experiments for each group). Monkey embryoids with clear embryonic disc-like structure including epiblast, amnion cavity and yolk sac cavity were calculated for statistical analysis. Each dot represents one repeat. k, Development efficiency of single cultured monkey embryoids from D17 to D25. Monkey embryoids with clear embryonic disc-like structure were calculated for statistical analysis (62 monkey embryoids on D17 from 3 independent experiments were used to track developmental efficiency until to D25). l, Measurement of long diameters of whole embryo and embryonic disc length of monkey embryoids from D13 to D25 respectively (n = 20 for D13, D15, D17, D20 and D23 groups; n = 16 for D25 group from 5 independent experiments). m, Representative 3D images showing the typical lateral view and dorsal view of monkey embryoids on D17 by light sheet microscope analysis after whole-mount IF staining of with TBXT (gray) and DAPI (Supplementary Video 14). TBXT, a marker for PS (primitive streak). Scale bars, 200 μm. n, Representative 3D images showing the typical lateral view of monkey embryoids on D15 by light sheet microscope analysis after whole-mount IF staining with OCT4 (gray), TFAP2A (red), and SOX17 (green) (Supplementary Video 15). OCT4, a marker for epiblast; TFAP2A, a marker for amnion cells; SOX17, a marker for yolk sac. Scale bars, 200 μm. c, e, j, l, Data are presented as mean ± SEM.
