Abstract：
An organoid can be defined as a three-dimensional organ-like structure formed from organ-specific progenitor cells. Organ progenitor cells were empirically found to self-organize three-dimensional tissues when they were aggregated and cultivated in vitro. While this nature power of progenitor cells has an amazing potential to recreate artificial organs in vitro, there had been difficulty to apply this technology to human organs due to the inaccessibility to human progenitor cells until human-induced pluripotent stem cell (hiPSC) was invented by Takahashi and Yamanaka in 2007. As embryonic stem cells do, hiPSCs also have pluripotency to give rise to any organs/tissues cell types, including the kidney, via directed differentiation. Here, we provide a detailed protocol for generating kidney organoids using human pluripotent stem cells. The protocol differentiates human pluripotent stem cells into the posterior primitive streak. This is followed by the simultaneous induction of posterior and anterior intermediate mesoderm that are subsequently aggregated and undergo self-organization into the kidney organoid. Such kidney organoids are comprised of all anticipated kidney cell types including nephrons segmented into the glomerulus, proximal tubule, loop of Henle, and distal tubule as well as the collecting duct, endothelial network, and renal interstitium.
摘要：
类器官可以定义为由器官特异性祖细胞形成的三维器官样结构。根据经验发现，器官祖细胞在体外聚集和培养时会自组织三维组织。虽然祖细胞的这种自然力量具有在体外重建人造器官的惊人潜力，在高桥和山中在2007年发明人类诱导多能干细胞（hiPSC）之前，由于人类祖细胞的难以接近性，将这项技术应用于人体器官一直存在困难。就像胚胎干细胞一样，hiPSC还具有多能性，以产生任何器官/组织细胞类型，包括肾脏,通过定向分化。这里,我们提供了使用人多能干细胞产生肾脏类器官的详细方案.该方案将人多能干细胞分化为后原始条纹。随后同时诱导后中间中胚层和前中间中胚层，这些中胚层随后聚集并自组织到肾脏类器官中。此类肾脏类器官由所有预期的肾脏细胞类型组成，包括分段为肾小球的肾单位，近端小管，Henle的循环,和远端小管以及收集管，内皮网络，和肾间质.
