Fig. 5

Antiapoptosis enables interspecies chimera with naive hPSCs. a Phase contrast of 3i and 5i naïve cells converted from primed UH10-DsRed hiPSCs; scale bars, 100 µm. b RT-qPCR analysis of BMI1 expression in the indicated cells; mean + SEM of three independent samples, ***p < 0.001. c Alkaline phosphatase staining on colonies formed by the indicated cells after plating of 50,000 cells on feeder for 7 days. d Statistics of the numbers of the indicated DsRed + cells in injected mouse blastocysts after injection of the ten indicated DsRed + cells in later morulas or early blastocysts and 1-day culture in vitro; scale bars, 50 µm; mean ± SEM of 40 (3i) or 37 (5i) samples, ***p < 0.001. e Representative fluorescence images of mouse embryos stained with Annexin V after injection of ten indicated DsRed⁺ cells in later morulas or early blastocysts and 1-day culture in vitro. White arrow, DsRed⁺/Annexin V⁺ cells; Yellow arrow, DsRed⁺/Annexin V⁻ cells. Scale bars, 20 µm. f Statistics of the percentage of the engrafted DsRed+ cells with Annexin V+ in the mouse blastocysts; mean + SEM of 40 (3i) or 37 (5i) samples, ***p < 0.001. g Summary of chimera assays with injection of ten indicated DsRed⁺ cells at the 8C stage embryo, and followed 48–60 h in vitro development into blastocyst stage. h Representative images showing the integration of indicated cells coexpressed OCT4, ICM marker or CDX2, early trophoblast marker in cultured chimeric embryos. White arrow, DsRed⁺/OCT4⁺ cells; scale bars, 20 µm. i Representative quantitative genomic PCR analysis of the human mitochondria DNA in E10.5 mouse embryos after injection of the indicated cells. A human DNA control (H), a mouse DNA control (M) and a series of human−mouse cell dilutions (1/100 to 1/1000,000) were run in parallel to estimate the degree of human cell integration. The dashed line indicates the detection level of one human cell in 10,000 mouse cells. j Summary of chimera assays of the indicated cells injectied at the later morula or early blastocyst stage