Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China; Beijing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China.
Colorado Center for Reproductive Medicine, Lone Tree, CO 80124, USA.
Cell Stem Cell. 2023 Sep 7;30(9):1246-1261.e9. doi: 10.1016/j.stem.2023.08.002.
Recent advances in human blastoids have opened new avenues for modeling early human development and implantation. One limitation of our first protocol for human blastoid generation was relatively low efficiency. We now report an optimized protocol for the efficient generation of large quantities of high-fidelity human blastoids from naive pluripotent stem cells. This enabled proteomics analysis that identified phosphosite-specific signatures potentially involved in the derivation and/or maintenance of the signaling states in human blastoids. Additionally, we uncovered endometrial stromal effects in promoting trophoblast cell survival, proliferation, and syncytialization during co-culture with blastoids and blastocysts. Side-by-side single-cell RNA sequencing revealed similarities and differences in transcriptome profiles between pre-implantation blastoids and blastocysts, as well as post-implantation cultures, and uncovered a population resembling early migratory trophoblasts during co-culture with endometrial stromal cells. Our optimized protocol will facilitate broader use of human blastoids as an accessible, perturbable, scalable, and tractable model for human blastocysts.
人类类胚体的最新进展为模拟早期人类发育和着床开辟了新途径。我们之前生成人类类胚体的方案存在一个局限性，那就是效率相对较低。现在，我们报告了一种优化的方案，可以从原始多能干细胞高效生成大量高保真的人类类胚体。该方案可进行蛋白质组学分析，鉴定出可能参与人类类胚体信号状态的衍生和/或维持的磷酸化特异性特征。此外，我们还发现子宫内膜基质细胞在与类胚体和囊胚共培养时，能够促进滋养层细胞的存活、增殖和融合。单细胞 RNA 测序的平行分析揭示了类胚体与囊胚以及着床后培养物在转录组谱方面的相似性和差异，并在与子宫内膜基质细胞共培养时发现了一个类似于早期迁移滋养层细胞的细胞群。我们的优化方案将促进更广泛地使用人类类胚体作为一种易于接近、可干扰、可扩展和可操作的人类囊胚模型。
