Fig. 1: Optimizing human naive ES cell differentiation towards extra-embryonic lineages competent for early post-implantation SEM generation.

a, Scheme of the tested induction (iGATA4, iGATA6, iCDX2 and iGATA3) and media conditions for generating the three different extra-embryonic lineages constituting the post-implantation human embryo (right) from HENSM naive PS cells (nPS cells). Epiblast (cyan), hypoblast (yellow), ExEM (grey) and trophoblast (magenta) compartments. b, Schematic (top) and FACS plots (bottom) of PDGFRA versus SSC for PrE-like and ExEM-like (PrE/ExEM) cell induction using iGATA4 with doxycycline (DOX) in HENSM after 6 days (right) and the control condition of naive cells (WT cells without iGATA4, left). c, Scheme (top) and FACS plots (bottom) of PDGFRA versus SSC for PrE/ExEM-like cell induction using iGATA4 and iGATA6 with DOX for 6 days in different media conditions as indicated (C10F4PDGF, RACL or RCL). d, Scheme (top) and FACS plots (bottom) of PDGFRA for PrE/ExEM-like cells using WT nES cells induced for 6 days in different media conditions (N2B27, RCL or RCL for 3 days followed by 3 days of N2B27). e, Immunofluorescence images of WT nES cells (WIBR3 line) induced for 6 days in RCL medium for SOX17, BST2 and nuclei (DAPI). Outline indicates mutually exclusive expression pattern of SOX17 and BST2. f, Scheme (top) and FACS plots (bottom) of ENPEP against TACSTD2 for TE-like lineage induction of HENSM nES cells (WIBR3 line) using the BAP(J) regimen for 3 days. g, Immunofluorescence images of day 6 SEM aggregates stained for OCT4, SOX17 and SDC1. The TE regimens used for SEM are indicated. Scale bars, 100 µm.