Shanghai Key Laboratory of Maternal Fetal Medicine, Clinical and Translational Research Center of Shanghai First Maternity and Infant Hospital, Frontier Science Center for Stem Cell Research, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
Translational Medical Center for Stem Cell Therapy and Institute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
Protein Cell. 2023 Apr 21;14(4):262-278. doi: 10.1093/procel/pwac029.
Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
自发形成的胚状体来源于扩展多能干细胞 (EPS)，在研究植入后胚胎发育和相关疾病方面具有巨大的潜力。然而，EPS 胚状体的植入后发育能力有限，限制了其进一步的应用。在这项研究中，单细胞转录组分析表明，EPS 胚状体的“滋养外胚层 (TE)-样结构”主要由原始内胚层 (PrE)-相关细胞组成，而不是 TE-相关细胞。我们进一步在 EPS 细胞培养中鉴定出 PrE 样细胞，这些细胞有助于形成具有 TE 样结构的胚状体。通过抑制 MEK 信号或敲除 EPS 细胞中的 Gata6 抑制 PrE 细胞分化，显著抑制了 EPS 胚状体的形成。此外，我们证明了通过将 EPS 衍生的双谱系胚胎样结构 (BLES)与四倍体胚胎或四倍体 TE 细胞组合，可以重建胚泡样结构，这些结构可以正常植入并发育成活胎。总之，我们的研究表明，TE 的改善对于体外使用干细胞构建功能性胚胎至关重要。
