European Molecular Biology Laboratory (EMBL), Barcelona, Barcelona, Spain.
EMBL Heidelberg, Developmental Biology Unit, Heidelberg, Germany.
Methods Mol Biol. 2021;2258:131-147. doi: 10.1007/978-1-0716-1174-6_10.
Gastruloids are embryonic organoids made from small, defined numbers of mouse embryonic stem cells (mESCs) aggregated in suspension culture, which over time form 3D structures that mimic many of the features of early mammalian development. Unlike embryoid bodies that are usually disorganized when grown over several days, gastruloids display distinct, well-organized gene expression domains demarcating the emergence of the three body axes, anteroposterior axial elongation, and implementation of collinear Hox transcriptional patterns over 5-7 days of culture. As such gastruloids represent a useful experimental system that is complementary to in vivo approaches in studying early developmental patterning mechanisms regulating the acquisition of cell fates. In this protocol, we describe the most recent method for generating gastruloids with high reproducibility, and provide a comprehensive list of possible challenges as well as steps for protocol optimization.
类囊胚是从小鼠胚胎干细胞（mESCs）中聚集在悬浮培养中的小而定义数量的细胞制成的胚胎类器官，随着时间的推移形成模拟早期哺乳动物发育的许多特征的 3D 结构。与通常在几天内生长时无组织的胚状体不同，类囊胚显示出明显的、组织良好的基因表达域，标志着三个体轴的出现、前后轴向伸长以及在 5-7 天的培养过程中实现共线性 Hox 转录模式。因此，类囊胚代表了一种有用的实验系统，与体内方法相辅相成，可用于研究调节细胞命运获得的早期发育模式形成机制。在本方案中，我们描述了一种具有高重复性的生成类囊胚的最新方法，并提供了可能出现的挑战以及优化方案步骤的综合清单。
