多能干细胞衍生的肾脏类器官作为终末期肾病患者的潜在辅助移植组织以及作为研究肾脏疾病和药物发现的平台具有广阔的前景。为了建立准确的模型，彻底表征这些类器官内细胞成分的形态特征和成熟阶段至关重要。肾单位是肾脏的功能单位，具有与其特定功能直接相关的独特形态结构。高空间分辨率成像作为一种捕获超微结构细节的强大技术而出现，而这些细节可能会被免疫荧光成像和 scRNA 测序等其他方法所忽视。在我们的研究中，我们应用了能够无缝缝合电子显微镜生成的虚拟载玻片的软件，从而获得组织载玻片的高清概览。通过这项技术，我们可以全面表征移植到小鼠肾囊下的肾脏类器官的发育和成熟。这些类器官在移植后表现出先进的超微结构发育，包括肾小体中滤过屏障的形成、近端小管中微绒毛的存在以及连接小管中类似于成人肾脏中所见的各种类型的细胞亚分割。这种超微结构特征为了解肾类器官内肾单位节段的结构发育和功能形态以及如何通过移植等干预措施来推进它们提供了宝贵的见解。研究亮点 高分辨率成像对于确定 hiPSC 来源的肾脏类器官的形态成熟至关重要。移植后，观察到肾单位节段的精细超微结构发育，例如肾小球滤过屏障的发育。© 2023 作者。显微镜研究和技术由 Wiley periodicals LLC 出版。

Pluripotent stem cell-derived kidney organoids hold great promise as a potential auxiliary transplant tissue for individuals with end-stage renal disease and as a platform for studying kidney diseases and drug discovery. To establish accurate models, it is crucial to thoroughly characterize the morphological features and maturation stages of the cellular components within these organoids. Nephrons, the functional units of the kidney, possess distinct morphological structures that directly correlate with their specific functions. High spatial resolution imaging emerges as a powerful technique for capturing ultrastructural details that may go unnoticed with other methods such as immunofluorescent imaging and scRNA sequencing. In our study, we have applied software capable of seamlessly stitching virtual slides generated from electron microscopy, resulting in high-definition overviews of tissue slides. With this technology, we can comprehensively characterize the development and maturation of kidney organoids when transplanted under the renal capsule of mice. These organoids exhibit advanced ultrastructural developments upon transplantation, including the formation of the filtration barrier in the renal corpuscle, the presence of microvilli in the proximal tubule, and various types of cell sub-segmentation in the connecting tubule similarly to those seen in the adult kidney. Such ultrastructural characterization provides invaluable insights into the structural development and functional morphology of nephron segments within kidney organoids and how to advance them by interventions such as a transplantation. Research Highlights High-resolution imaging is crucial to determine morphological maturation of hiPSC-derived kidney organoids. Upon transplantation, refined ultrastructural development of nephron segments was observed, such as the development of the glomerular filtration barrier.© 2023 The Authors. Microscopy Research and Technique published by Wiley Periodicals LLC.