Metabolically reprogrammed blastoids from totipotent-like stem cells exhibit enhanced tissue-specific organ-like development and placental formation

While blastoids derived from pluripotent stem cells recapitulate pre- and early post-implantation embryogenesis, their long-term post-implantation development remains constrained. We define Snαnv, a chemical cocktail that promotes oxidative-phosphorylation-driven metabolic reprogramming, to efficiently generate ESC-blastoids with proper trophectoderm (TE)/inner cell mass (ICM) from conventional mouse ESCs at an efficiency of approximately 70%. These blastoids closely resemble blastocysts in morphology and cell-lineage allocation, with high Cdx2 expression in TE-like cells and notably display similar metabolic profiles. Furthermore, enhancing ESC totipotency via pladienolide B (PlaB) and crotonic acid (CA) augments the developmental potential of resulting PC-blastoids, which yield gonad-like tissues at E10.5, beating heart-like structures, and blood vessels under specific in vitro induction. These metabolically reprogrammed PC-blastoids are transcriptionally analogous to blastocysts by scRNA-seq. Moreover, they generate E7.0/E7.5-like embryos and placental structures comparable to E8.5 counterparts in vivo. Together, mimicking blastocyst metabolism and enhancing ESCs' totipotency yield blastoids with notably improved developmental competence. Here they show that metabolic reprogramming combined with enhanced totipotency confers blastoids with improved competence, generating E10.5 gonad-like tissues and beating heart-like structures in vitro and enhancing placental development in vivo.