Creating complex multicellular kidney organoids from pluripotent stem cells shows great promise. Further improvements in differentiation outcomes, patterning, and maturation of specific cell types are, however, intrinsically limited by standard tissue culture approaches. We describe a novel full factorial microbioreactor array-based methodology to achieve rapid interrogation and optimization of this complex multicellular differentiation process in a facile manner. We successfully recapitulate early kidney tissue patterning events, exploring more than 1000 unique conditions in an unbiased and quantitative manner, and define new media combinations that achieve near-pure renal cell type specification. Single-cell resolution identification of distinct renal cell types within multilayered kidney organoids, coupled with multivariate analysis, defined the definitive roles of Wnt, fibroblast growth factor, and bone morphogenetic protein signaling in their specification, exposed retinoic acid as a minimal effector of nephron patterning, and highlighted critical contributions of induced paracrine factors on cell specification and patterning.

中文翻译：

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人多能细胞在灌注下的多变量模式揭示了诱导的旁分泌因子在肾脏类器官发育中的关键作用。

从多能干细胞创建复杂的多细胞肾脏类器官显示出巨大的希望。但是，特定组织类型的分化结果，模式和成熟的进一步改善本质上受到标准组织培养方法的限制。我们描述了一种新颖的基于全因子微生物反应器阵列的方法，以快速的方式来实现对这种复杂的多细胞分化过程的快速询问和优化。我们成功地概括了早期肾脏组织的模式事件，以无偏见和定量的方式探索了1000多种独特条件，并定义了可实现近乎纯净的肾细胞类型规范的新培养基组合。多层肾脏类器官内单个肾细胞类型的单细胞分辨率鉴定，以及多变量分析，