The study of brain development in humans is limited by the lack of tissue samples and suitable in vitro models. Here, we model early human neural tube development using human embryonic stem cells cultured in a microfluidic device. The approach, named microfluidic-controlled stem cell regionalization (MiSTR), exposes pluripotent stem cells to signaling gradients that mimic developmental patterning. Using a WNT-activating gradient, we generated a neural tissue exhibiting progressive caudalization from forebrain to midbrain to hindbrain, including formation of isthmic organizer characteristics. Single-cell transcriptomics revealed that rostro-caudal organization was already established at 24 h of differentiation, and that the first markers of a neural-specific transcription program emerged in the rostral cells at 48 h. The transcriptomic hallmarks of rostro-caudal organization recapitulated gene expression patterns of the early rostro-caudal neural plate in mouse embryos. Thus, MiSTR will facilitate research on the factors and processes underlying rostro-caudal neural tube patterning.

缺乏组织样本和合适的体外模型限制了人类大脑发育的研究。在这里，我们使用在微流体设备中培养的人类胚胎干细胞对人类神经管的早期发育进行建模。该方法名为微流控干细胞区域化（MiSTR），将多能干细胞暴露于模仿发育模式的信号梯度中。使用WNT激活梯度，我们生成了神经组织，该神经组织表现出从前脑到中脑再到后脑的渐进钙化，包括形成峡部组织者特征。单细胞转录组学表明，在分化的24小时已经建立了尾状尾尾组织，并且神经特异性转录程序的第一个标记在48小时出现在了尾状细胞中。ros尾尾组织的转录组特征概括了小鼠胚胎中早期尾尾神经板的基因表达模式。因此，MiSTR将促进对猪尾神经管构图的因素和过程的研究。