Abstract
Mammalian embryogenesis begins with a totipotent zygote. Blastocyst-like structures can be captured by aggregated cells with extended pluripotent properties in a three-dimensional (3D) culture system. However, the efficiency of generating blastoids is low, and it remains unclear whether other reported totipotent-like stem cells retain a similar capacity. In this study, we demonstrated that spliceosomal repression-induced totipotent blastomere-like cells (TBLCs) form blastocyst-like structures within around 80% of all microwells. In addition, we generated blastoids initiating from a single TBLC. TBLC-blastoids express specific markers of constituent cell lineages of a blastocyst and resemble blastocyst in cell-lineage allocation. Moreover, single-cell RNA sequencing revealed that TBLC-blastoids share a similar transcriptional profile to natural embryos, albeit composed of fewer primitive endoderm-like cells. Furthermore, TBLC-blastoids can develop beyond the implantation stage in vitro and induce decidualization in vivo. In summary, our findings provided an alternative cell type to efficiently generate blastoids for the study of early mouse embryogenesis.
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Data availability
All raw sequencing data can be accessed at the NCBI Gene Expression Omnibus (GEO) (accession number: GSE197779). The raw count of scRNA-sequencing data of E3.5–E7.5 embryos (in-house data) can be found in the Supplementary File.
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Acknowledgements
We thank Dr. Peng Du (Peking University) for their TC1-EGFP ESCs and TC1-EGFP TBLCs, thank Dr. Jiekai Chen and Dr. Duanqing Pei for their single-cell RNA-seq data of blastocysts, and thank Dr. Xudong Fu (Zhejiang University) for his helpful comments on this manuscript. This work was supported by the National Natural Science Foundation of China (32070800).
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Zhang, P., Zhai, X., Huang, B. et al. Highly efficient generation of blastocyst-like structures from spliceosomes-repressed mouse totipotent blastomere-like cells. Sci. China Life Sci. 66, 423–435 (2023). https://doi.org/10.1007/s11427-022-2209-3
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DOI: https://doi.org/10.1007/s11427-022-2209-3