Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells.
Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells.
复制标题
人类多能干细胞的肾单位祖细胞和肾脏器官的产生。
DOI：
10.1038/nprot.2016.170
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发表时间：
2017-01
期刊：
影响因子：
14.8
通讯作者：
Bonventre JV
中科院分区：
文献类型：
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作者：
Morizane R;Bonventre JV
已经开发出多种方案，证明其能够将人类多能干细胞 (hPSC) 分化为肾脏结构。我们的目标是开发一种高效的方案来生成肾单位祖细胞 (NPC) 和肾脏类器官，以促进组织工程、疾病建模和化学筛选的应用。在这里，我们描述了一个详细的方案，可在 hPSC 分化后 9 天内高效生产 NPC（80-90%）。使用适合化学筛选的 96 孔板，在 12 天内以高重现性从 NPC 生成肾脏类器官。该协议需要培养 hPSC 的技能，并仔细注意指示分化的形态变化。该肾脏类器官系统为研究人类肾脏发育、肾脏疾病建模、肾毒性和肾脏再生提供了一个平台。该系统为在适当的肾单位和基质背景下使用分化的人类细胞进行细胞内和肾脏间室相互作用的体外研究提供了模型。
A variety of protocols have been developed which demonstrate a capability to differentiate human pluripotent stem cells (hPSCs) into kidney structures. Our goal was to develop a high efficiency protocol to generate nephron progenitor cells (NPCs) and kidney organoids to facilitate applications for tissue engineering, disease modeling and chemical screening. Here, we describe a detailed protocol resulting in high efficiency production (80–90%) of NPCs within 9 days of differentiation from hPSCs. Kidney organoids were generated from NPCs within 12 days with high reproducibility using 96-well plates suitable for chemical screening. The protocol requires skills in culturing hPSCs and careful attention to morphological changes indicative of differentiation. This kidney organoid system provides a platform for studies of human kidney development, modeling of kidney diseases, nephrotoxicity, and kidney regeneration. The system provides a model for in vitro study of intracellular and kidney inter-compartmental interactions using differentiated human cells in an appropriate nephron and stromal context.