Chen Bohan, Khan Hina, Yu Zhiyuan, Yao LiAng, Freeburne Emily, Jo Kyoung, Johnson Craig, Heemskerk Idse
Department of Computational Medicine and Bioinformatics, University of Michigan Medical School, Ann Arbor, MI, USA.
Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USA.
Nat Methods. 2025 May 7. doi: 10.1038/s41592-025-02669-4.
Micropatterned human pluripotent stem cells treated with BMP4 (two-dimensional (2D) gastruloids) are among the most widely used stem cell models for human gastrulation. Due to its simplicity and reproducibility, this system is ideal for high-throughput quantitative studies of tissue patterning and has led to many insights into the mechanisms of mammalian gastrulation. However, 2D gastruloids have been studied only up to about 2 days owing to a loss of organization beyond this time with earlier protocols. Here we report an extended 2D gastruloid model to up to 10 days. We discovered a phase of highly reproducible morphogenesis between 2 and 4 days during which directed migration from the primitive streak-like region gives rise to a mesodermal layer beneath an epiblast-like layer. Multiple types of mesoderm arise with striking spatial organization including lateral plate mesoderm-like cells on the colony border and paraxial mesoderm-like cells further inside the colony. Single-cell transcriptomics showed strong similarity of these cells to mesoderm in human and nonhuman primate embryos. Our results illustrate that extended culture of 2D gastruloids provides a powerful model for human mesoderm differentiation and morphogenesis.
用骨形态发生蛋白4（BMP4）处理的微图案化人类多能干细胞（二维（2D）类原肠胚）是用于人类原肠胚形成研究的最广泛使用的干细胞模型之一。由于其简单性和可重复性，该系统非常适合用于组织图案化的高通量定量研究，并为哺乳动物原肠胚形成机制带来了许多见解。然而，由于早期方案在超过这个时间后组织会失去结构，二维类原肠胚仅被研究到大约2天。在这里，我们报告了一种可延长至10天的二维类原肠胚模型。我们发现在2至4天之间存在一个高度可重复的形态发生阶段，在此期间，从原始条纹样区域的定向迁移会在类似上胚层的层下方形成一个中胚层。多种类型的中胚层以显著的空间组织形式出现，包括集落边界上的侧板中胚层样细胞和集落内部更深处的近轴中胚层样细胞。单细胞转录组学显示这些细胞与人类和非人类灵长类胚胎中的中胚层具有很强的相似性。我们的结果表明，二维类原肠胚的延长培养为人类中胚层分化和形态发生提供了一个强大的模型。
