Figure 3.
BMEx overlay differentiation promotes differentiation to hPGCLCs alongside amniotic ectoderm- and mesoderm-like cells
(A) Uniform manifold approximation and projection (UMAP) plots showing cluster identification (ID) (left) and time period in days (right) using single-cell transcriptomics of several hPSC lines undergoing differentiation with BMEx overlay.
(B) Heatmap showing expression levels of the top 12 DEGs of each cluster.
(C) Expression of signature genes of cell types of interest on the UMAP plot from (A).
(D) UMAP showing integrated single-cell transcriptomics data from EB differentiation method (UCLA2 from Chen et al.11) and BMEx overlay method, highlighting the cells of EB differentiation method (left) and the BMEx overlay method (right).
(E) Expression of signature genes of cell types of interest on the UMAP plot from (D).
(F) Immunofluorescence for TFAP2A, TFAP2C, and GATA6 (top left); GATA3, SOX17, and TFAP2C (top right); and TFAP2A, GATA6, and PDGFRA (bottom) at day 5 with BMEx overlay. In the top panels, the dashed box is magnified (right), showing separated channels. The bottom panels depict a maximum intensity projection (MIP) and render image with a digital cross section showing separated channels (bottom part). Scale bars: 50 μm.
(G) Whole-mount immunofluorescence for TFAP2A, GATA6, and PDGFRA in human WG9 amnion. In the top panels, a digital cross section (right) shows separated channels. The bottom panels depict a MIP and render image, also shown from the side (bottom). Scale bars: 50 μm. AELC, amniotic ectoderm-like cell; AMLC, amniotic mesoderm-like cell; iMeLCs, incipient mesoderm-like cells; PELC, primitive endoderm-like cell; Progen, progenitor cells.
See also Figure S2.