Single-cell RNA-seq for mouse NrESC-blastoids

Embryonic stem cells upon extrinsic induction could self-assemble into blastocyst-like structures. However, the intrinsic regulation of such blastoid forming potential remain to be addressed. We discover that the activity of nuclear receptor subfamily 1, group H, member 2 (Nr1h2) in expanded potential stem cell (EPSC) positively correlates with blastoid efficiency and quality. In addition, Nr1h2 agonist, T0901317, improves natural blastocyst development. Surprisingly, Nr1h2-activated ESC (NrESC) is rewired towards a distinct pluripotency state that is capable of self-organizing into blastoids and contribute to embryonic and extraembryonic lineage. We aim at characterizing the NrESC-derived blastoids by single-cell RNA-seq and show that blastoid comprise of epiblast-like, trophectoderm-like and primitive endoderm-like populations, which is the same as blastocyst composition and shows respective gene marker expression for each cluster. Thus, NrESC has attained a unique expanded pluripotency state driven by Nr1h2 activation. Overall design: Single-cell RNA-seq was applied on mouse NrESC-derived blastoids to examine the single cell transcriptome of different lineages in the organoid