Yilmaz Alperen, Gurhan Gulben, Comar Mehmet-Yunus, Viukov Sergey, Serfaty Inbal, Gayretli Mert, Golenchenko Sergey, Lokshtanov Dmitry, Ashouokhi Shahd, Polanco Angel, Berlad Idan, Ha Tae-Won, Aguilera-Castrejon Alejandro, Tarazi Shadi, Cohen Marina, Livnat Nir, Kumar Komal, Cholakkal Hisham, Levy Nathan, Yosef Nir, Khatib Nizar, Kakun Reli Rachel, Kedmi Merav, Nachman Inbal Bolocan, Keren-Shaul Hadas, Addadi Yoseph, Orenbuch Ayelet-Hashahar, Korovin Karina, Molchadsky Alina, Hochedlinger Konrad, Gafni Ohad, Maza Itay, Novershtern Noa, Oldak Bernardo, Hanna Jacob H
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
Cell Stem Cell. 2025 Aug 7. doi: 10.1016/j.stem.2025.07.005.
The generation of post-gastrulation stem cell-derived mouse embryo models (SEMs) exclusively from naive embryonic stem cells (nESCs) has underscored their ability to give rise to embryonic and extra-embryonic lineages. However, existing protocols for mouse SEMs rely on the separate induction of extra-embryonic lineages and on ectopic expression of transcription factors to induce nESC differentiation into trophectoderm (TE) or primitive endoderm (PrE). Here, we demonstrate that mouse nESCs and naive induced pluripotent stem cells (niPSCs) can be simultaneously co-induced, via signaling pathway modulation, to generate PrE and TE extra-embryonic cells that self-organize into embryonic day (E) 8.5-E8.75 transgene-free (TF) SEMs. We also devised an alternative condition (AC) naive media that in vitro stabilizes TF-SEM-competent OCT4+/NANOG+ nESC colonies that co-express antagonistic CDX2 and/or GATA6 extra-embryonic fate master regulators and self-renew while remaining poised for TE and PrE differentiation, respectively. These findings improve mouse SEM strategies and shed light on amplifying an inherent and dormant extra-embryonic plasticity of mouse naive pluripotent cells in vitro.
仅从原始胚胎干细胞（nESCs）生成原肠胚形成后干细胞衍生的小鼠胚胎模型（SEMs），突出了它们产生胚胎和胚外谱系的能力。然而，现有的小鼠SEMs方案依赖于胚外谱系的单独诱导以及转录因子的异位表达，以诱导nESCs分化为滋养外胚层（TE）或原始内胚层（PrE）。在这里，我们证明，通过信号通路调节，可以同时共诱导小鼠nESCs和原始诱导多能干细胞（niPSCs），以产生PrE和TE胚外细胞，这些细胞自组装成无转基因（TF）的胚胎第8.5天至8.75天的SEMs。我们还设计了一种替代条件（AC）原始培养基，该培养基在体外稳定具有TF-SEM能力的OCT4+/NESC集落，这些集落共表达拮抗的CDX2和/或GATA6胚外命运主调节因子，并自我更新，同时分别保持对TE和PrE分化的准备状态。这些发现改进了小鼠SEMs策略，并为在体外放大小鼠原始多能细胞固有的、休眠的胚外可塑性提供了线索。
