Gantner Carlos W, Weatherbee Bailey A T, Wang Yuntao, Zernicka-Goetz Magdalena
Mammalian Embryo and Stem Cell Group, Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
Stem Cell Embryo Models Group, Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.
Nat Protoc. 2025 Jan;20(1):67-91. doi: 10.1038/s41596-024-01042-7. Epub 2024 Sep 11.
The embryonic and extraembryonic tissue interactions underlying human embryogenesis at implantation stages are not currently understood. We have generated a pluripotent stem cell-derived model that mimics aspects of peri-implantation development, allowing tractable experimentation otherwise impossible in the human embryo. Activation of the extraembryonic lineage-specific transcription factors GATA6 and SOX17 (hypoblast factors) or GATA3 and TFAP2C (encoding AP2γ; trophoblast factors) in human embryonic stem (ES) cells drive conversion to extraembryonic-like cells. When combined with wild-type ES cells, self-organized embryo-like structures form in the absence of exogenous factors, termed human inducible embryoids (hiEmbryoids). The epiblast-like domain of hiEmbryoids polarizes and differentiates in response to extraembryonic-secreted extracellular matrix and morphogen cues. Extraembryonic mesenchyme, amnion and primordial germ cells are specified in hiEmbryoids in a stepwise fashion. After establishing stable inducible ES lines and converting ES cells to RSeT culture media, the protocol takes 7-10 d to generate hiEmbryoids. Generation of hiEmbryoids can be performed by researchers with basic expertise in stem cell culture.
目前尚不清楚人类胚胎着床阶段胚胎发育和胚外组织相互作用的机制。我们构建了一种多能干细胞衍生模型，该模型模拟了着床周围发育的各个方面，使得原本在人类胚胎中无法进行的易于处理的实验成为可能。在人类胚胎干细胞（ES细胞）中激活胚外谱系特异性转录因子GATA6和SOX17（内胚层因子）或GATA3和TFAP2C（编码AP2γ；滋养层因子）可驱动其转化为类胚外细胞。当与野生型ES细胞结合时，在没有外源性因子的情况下会形成自组织的胚胎样结构，称为人类诱导胚状体（hiEmbryoids）。hiEmbryoids的上胚层样结构域会响应胚外分泌的细胞外基质和形态发生素信号而极化并分化。胚外间充质、羊膜和原始生殖细胞在hiEmbryoids中逐步形成。在建立稳定的诱导ES细胞系并将ES细胞转换为RSeT培养基后，该方案需要7-10天来生成hiEmbryoids。具有干细胞培养基本专业知识的研究人员即可进行hiEmbryoids的生成。
