Abstract：
Our understanding in the inherent properties of human pluripotent stem cells (hPSCs) have made possible the development of differentiation procedures to generate three-dimensional tissue-like cultures, so-called organoids. Here we detail a stepwise methodology to generate kidney organoids from hPSCs. This is achieved through direct differentiation of hPSCs in two-dimensional monolayer culture toward the posterior primitive streak fate, followed by induction of intermediate mesoderm-committed cells, which are further aggregated and cultured in three-dimensions to generate kidney organoids containing segmented nephron-like structures in a process that lasts 20 days. We also provide a concise description on how to assess renal commitment during the time course of kidney organoid generation. This includes the use of flow cytometry and immunocytochemistry analyses for the detection of specific renal differentiation markers.
摘要：
我们对人类多能干细胞（hPSC）固有特性的理解使分化程序的发展成为可能，以产生三维组织样培养物。所谓的类器官。在这里，我们详细介绍了从hPSC产生肾类器官的逐步方法。这是通过在二维单层培养中hPSC向后原始条纹命运的直接分化来实现的。然后诱导中间中胚层定向细胞，在持续20天的过程中进一步聚集和三维培养以产生含有分段肾单位样结构的肾脏类器官。我们还提供了有关如何在肾脏类器官生成过程中评估肾脏承诺的简要描述。这包括使用流式细胞术和免疫细胞化学分析来检测特定的肾分化标志物。
