Mouse totipotent stem cells captured and maintained through spliceosomal repression

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 ESC...

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Bibliographic Details
Published in:Cell 2021-05, Vol.184 (11), p.2843-2859.e20
Main Authors: Shen, Hui, Yang, Min, Li, Shiyu, Zhang, Jing, Peng, Bing, Wang, Chunhui, Chang, Zai, Ong, Jennie, Du, Peng
Format: Article
Language:English
Subjects:
Animals
Blastomeres - cytology
Cell Differentiation - genetics
Cell Line
Cell Lineage - genetics
chimeras
Embryo, Mammalian - cytology
embryonic
embryonic stem cell
Embryonic Stem Cells - cytology
extraembryonic
Female
Male
Mice
Mice, Inbred C57BL
Mice, Inbred ICR
Mouse Embryonic Stem Cells - cytology
pladienolide B
pluripotent
spliceosome
splicing
totipotent
Totipotent Stem Cells - cytology
Totipotent Stem Cells - metabolism
Totipotent Stem Cells - physiology
transcriptome
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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. [Display omitted] •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. Totipotent blastomere-like cells are reprogrammed from mouse pluripotent stem cells and cultured stably for multiple passages in vitro by spliceosomal repression.
ISSN:0092-8674
1097-4172
DOI:10.1016/j.cell.2021.04.020