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. Author manuscript; available in PMC: 2020 Oct 17.
Published in final edited form as: Cell. 2019 Oct 17;179(3):687–702.e18. doi: 10.1016/j.cell.2019.09.029

Figure 5. In Vitro Developmental Potential of EPS-blastoids.

Figure 5.

(A) Phase contrast image of de novo derived ES cell lines from EPS-blastoids.

(B) Brightfield image of two littermates generated from blastocyst injected with EPS-blastoid-derived ES cells showing that these cells can contribute to chimeric mice. The star symbol denotes a chimeric mouse.

(C) Phase contrast image of de novo derived TS cell lines from EPS-blastoids.

(D) Immunofluorescence staining of a placental section for CK8 and GFP. The panels below are the enlargement of the yellow boxed region. The placenta was delineated by a dotted line; dec, decidua layer; gc, giant cell layer; sp, spongiotrophoblast layer; laby, labyrinth layer.

(E) Phase contrast image of de novo derived XEN cell lines from EPS-blastoids.

(F) Brightfield image of a yolk sac overlaid with tdTomato epifluorescence image showing EPS-blastoid-derived XEN cells can contribute to the developing yolk sac. Td, tdTomato.

(G) Immunofluorescence staining of blastocyst-derived postimplantation embryo-like structures for TFAP2C and SOX2 (upper panel) or GATA6 and SOX2 (lower panel).

(H) Immunofluorescence staining of EPS-blastoid-derived postimplantation embryo-like structures for TFAP2C and SOX2 (upper panel) or GATA4 and OCT4 (lower panel).

(I) Quantification of the percentage of postimplantation embryo-like and malformed structures formed after in vitro culture of blastocysts and EPS-blastoids. n = 3 and 4 independent assays for blastocysts and EPS-blastoids, respectively.

(J) Immunofluorescence staining of an EPS-blastoid-derived peri-implantation embryo-like structure for F-actin and NANOG showing the formation of rosette EPI-like structure.

(K) Immunofluorescence staining of an EPS-blastoid-derived postimplantation embryo-like structure for aPKC and SOX2. Yellow arrowhead denotes the apical domain.

(L and M) Immunofluorescence staining of a blastocyst- (L) or an EPS-blastoid- (M) derived postimplantation embryo-like structure for PCX and SOX2 (L) or PCX and OCT4 (M). Yellow arrowheads denote the enrichment of PCX protein around the lumens in both the EPI and ExE-like structure. PCX, podocalyxin.

Data are represented as mean ± SEM. Scale bar, 500 μm (D, upper), 100 μm (A, C, E, and F), 50 μm (G, H, K, L, and M), 20 μm (D, bottom; and J). Ho, Hoechst.

See also Figure S5.