Cell
Volume 184, Issue 11, 27 May 2021, Pages 2843-2859.e20
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Article
Mouse totipotent stem cells captured and maintained through spliceosomal repression

https://doi.org/10.1016/j.cell.2021.04.020Get rights and content
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Highlights

  • Spliceosomal repression reprograms pluripotent mouse ESCs to a totipotent state

  • PlaB enables in vitro culture of TBLCs molecularly close to totipotent blastomeres

  • TBLCs have robust bidirectional embryonic and extraembryonic differentiation potential

  • Pluripotent and totipotent genes respond differentially to spliceosomal repression.

Summary

Since establishment of the first embryonic stem cells (ESCs), in vitro culture of totipotent cells functionally and molecularly comparable with in vivo blastomeres with embryonic and extraembryonic developmental potential has been a challenge. Here we report that spliceosomal repression in mouse ESCs drives a pluripotent-to-totipotent state transition. Using the splicing inhibitor pladienolide B, we achieve stable in vitro culture of totipotent ESCs comparable at molecular levels with 2- and 4-cell blastomeres, which we call totipotent blastomere-like cells (TBLCs). Mouse chimeric assays combined with single-cell RNA sequencing (scRNA-seq) demonstrate that TBLCs have a robust bidirectional developmental capability to generate multiple embryonic and extraembryonic cell lineages. Mechanically, spliceosomal repression causes widespread splicing inhibition of pluripotent genes, whereas totipotent genes, which contain few short introns, are efficiently spliced and transcriptionally activated. Our study provides a means for capturing and maintaining totipotent stem cells.

Keywords

embryonic stem cell
totipotent
pluripotent
spliceosome
splicing
pladienolide B
transcriptome
embryonic
extraembryonic
chimeras

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