Morphogens choreograph the generation of remarkable cellular diversity in the developing nervous system. Differentiation of stem cells in vitro often relies upon the combinatorial modulation of these signaling pathways. However, the lack of a systematic approach to understand morphogen-directed differentiation has precluded the generation of many neural cell populations, and the general principles of regional specification and maturation remain incomplete. Here, we developed an arrayed screen of 14 morphogen modulators in human neural organoids cultured for over 70 days. Deconvolution of single-cell-multiplexed RNA sequencing data revealed design principles of brain region specification. We tuned neural subtype diversity to generate a tachykinin 3 (TAC3)-expressing striatal interneuron type within assembloids. To circumvent limitations of in vitro neuronal maturation, we used a neonatal rat transplantation strategy that enabled human Purkinje neurons to develop their hallmark complex dendritic branching. This comprehensive platform yields insights into the factors influencing stem cell-derived neural diversification and maturation.

中文翻译：

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在类器官中使用多重形态发生素筛选产生人类神经多样性


形态发生素在发育中的神经系统中编排了显着细胞多样性的产生。干细胞在 体外的分化通常依赖于这些信号通路的组合调节。然而，缺乏理解形态发生素定向分化的系统方法排除了许多神经细胞群的产生，并且区域规范和成熟的一般原则仍然不完整。在这里，我们在培养了 70 多天的人神经类器官中开发了 14 种形态发生调节剂的阵列筛选。单细胞多路复用 RNA 测序数据的反卷积揭示了大脑区域规范的设计原则。我们调整了神经亚型的多样性，在组装体内产生表达速激肽 3 （TAC3） 的纹状体中间神经元类型。为了规避体外神经元成熟的限制 ，我们使用了一种新生大鼠移植策略，使人类浦肯野神经元能够发展其标志性的复杂树突状分支。这个全面的平台可以深入了解影响干细胞衍生的神经多样化和成熟的因素。