GSE201747
Single-cell RNA sequencing analysis of mouse totipotent potential stem cells, extended pluripotent stem cells, embryonic stem cells, E17.5 TPS-derived chimeric placental cells, TPS-derived teratomas and TPS-derived blastoids
It is challenging to derive totipotent stem cells in vitro that functionally and molecularly resemble cells from totipotent embryos. Here, we report that a chemical cocktail enables the derivation of totipotent-like stem cells, designated as totipotent potential stem (TPS) cells, from 2-cell mouse embryos and extended pluripotent stem cells that can be stably maintained long-term in vitro. TPS cells shared features with 2-cell mouse embryos in terms of totipotency markers, transcriptome, chromatin accessibility and DNA methylation patterns. In vivo chimeric assays show that these cells have embryonic and extraembryonic developmental potentials at the single cell level. Moreover, TPS cells can be induced into blastocyst-like structures resembling preimplantation mouse blastocysts. Mechanistically, inhibition of HDAC1/2 and DOT1L activity and activation of RAR? signaling are important for inducing and maintaining totipotent features of TPS cells. Our study opens up a path toward fully capturing totipotent stem cells in vitro. Overall design: Total of 11 samples were analyzed, which include mouse totipotent potential stem (TPS) cells, extended pluripotent stem (EPS) cells, embryonic stem (ES) cells, E17.5 TPS-derived chimeric placental cells, TPS-derived teratomas and TPS-derived blastoids. For mouse TPS cells, 2 cell lines were analyzed, which included 1 cell line converted from mouse EPS cells and 1 cell line derived from 2-cell embryos. For mouse EPS cells, 2 cell lines were analyzed. For mouse ES cells, 2 cell lines were analyzed. For E17.5 TPS-derived chimeric placental cells, 2 samples were analyzed. For TPS-derived teratomas, 2 samples were analyzed. For TPS-derived blastoids, 1 sample was analyzed.