Handford Charlotte E, Panda Pallavi, Mohammad Choudhury Ikbal B, Amadei Gianluca, Zernicka-Goetz Magdalena
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2024;2767:123-134. doi: 10.1007/7651_2023_493.
In this chapter, we detail the experimental protocol leading to the generation of stem cell-based mouse embryo-like structures termed "ETiX-embryoids." ETiX-embryoids are formed from combined embryonic stem cells, trophoblast stem cells, and embryonic stem cells transiently induced to express Gata4. Cells are seeded into AggreWell dishes where they form aggregates that develop to resemble post-implantation mouse embryos following 4 days of culture. ETiX-embryoids establish an anterior signaling center and undergo gastrulation over the following 2 days. By day 7, ETiX-embryoids undergo neurulation and form an anterior-posterior axis with head folds at one end and a tail bud on the other. On day 8, they develop a brain and form a heart-like structure and a gut tube.
在本章中，我们详细介绍了生成名为“ETiX胚状体”的基于干细胞的小鼠胚胎样结构的实验方案。ETiX胚状体由组合的胚胎干细胞、滋养层干细胞以及短暂诱导表达Gata4的胚胎干细胞形成。将细胞接种到AggreWell培养皿中，它们在其中形成聚集体，经过4天培养后发育成类似植入后小鼠胚胎的结构。ETiX胚状体建立一个前部信号中心，并在接下来的2天内经历原肠胚形成。到第7天，ETiX胚状体进行神经胚形成，并形成前后轴，一端有头褶，另一端有尾芽。在第8天，它们发育出大脑，并形成类似心脏的结构和肠管。
