Extended culture of 2D gastruloids to model human mesoderm development.
Extended culture of 2D gastruloids to model human mesoderm development.
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二维原肠胚的扩展培养来模拟人类中胚层发育。
DOI：
10.1101/2024.03.21.585753
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发表时间：
2024
期刊：
影响因子：
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通讯作者：
Heemskerk,Idse
中科院分区：
文献类型：
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作者：
Chen,Bohan;Khan,Hina;Yu,Zhiyuan;Yao,LiAng;Freeburne,Emily;Jo,Kyoung;Johnson,Craig;Heemskerk,Idse
用BMP 4（二维（2D）类胃体）处理的微图案化的人多能干细胞是用于人原肠胚形成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，该系统是理想的高通量定量研究的组织图案，并导致了许多见解哺乳动物原肠胚形成的机制。然而，2D类胃窦仅被研究了约2天，因为早期方案在此时间之后失去了组织。在这里，我们报告了一个长达10天的扩展2D胃样模型。我们发现了一个阶段的高度可重复的形态发生之间的2和4天，在此期间，直接从原始条纹样区域的迁移引起下表胚样层的中胚层层。多种类型的中胚层具有显著的空间组织，包括在殖民地边界上的侧板中胚层样细胞和进一步在殖民地内的近轴中胚层样细胞。单细胞转录组学显示，这些细胞在人类和非人类灵长类动物胚胎中的中胚层具有很强的相似性。我们的研究结果表明，扩大培养的2D gastruloids提供了一个强大的模型，为人类中胚层分化和形态发生。
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.