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. 2022 Apr 14;17(5):1059–1069. doi: 10.1016/j.stemcr.2022.03.009

Figure 2.

Figure 2

MYCN/BCL2 promotes the integration and proliferation of hPSCs in interspecies pre-implantation embryos

(A and B) Survival and proliferation of dsRed-labeled hPSCs in pre-implantation embryos of different species. dsRed + hPSCs (10 cells for mouse, 3–5 cells for pig, and 7 cells for rabbit) were injected into 8-cell (8C)-stage embryos of indicated species and were cultured for 3 days in vitro, and the cell number of each indicated cell line was calculated. Scale bars, 20 μm. Mouse: mean + SEM of 20 (+B) or 20 (+N + B) samples; pig: mean + SEM of 18 (+B) or 18 (+N + B) samples; rabbit: mean + SEM of 16 (+B) or 16 (+N + B) samples. ∗∗∗p < 0.001.

(C) Contribution of hPSCs to embryonic and extra-embryonic embryonic lineages in mouse chimeras. Representative images showing the integration of M/B-hPSCs co-expressing OCT4, an ICM marker, or CDX2, an early trophoblast marker in cultured chimeric embryos. Yellow arrow, dsRed+/OCT4+ cells; white arrow, dsRed+/CDX2+ cells; scale bars, 20 μm.

(D) Representative images showing the integration of M/B-hPSC co-staining with hN (human cell nucleus-specific antibody) and GATA3 (early extra-embryonic lineage marker) in cultured chimeric embryos. White arrow, hN+/GATA3+ cells; scale bars, 20 μm.

(E) Summary of chimera assays with injection of ten indicated dsRed+ cells at the 8C-stage embryo, followed by 48–60 h in vitro development into the blastocyst stage.