Ultrastructural characterization of maturing iPSC‐derived nephron structures upon transplantation

• Published in: Microscopy research and technique 2024-03, Vol.87 (3), p.495-505
• Main Authors: Wiersma, L. E., Avramut, M. C., Koster, A. J., Berg, C. W., Rabelink, T. J.
• Format: Article
• Language: English
• Subjects: Electron microscopy; Filtration; Functional morphology; iPSC‐derived kidney organoids; Kidney diseases; Kidney transplantation; Kidneys; Maturation; Medical imaging; Morphology; Nephrons; Organoids; Pluripotency; regenerative medicine; Segments; Spatial discrimination; Spatial resolution; Stem cells; Stitching; Structure-function relationships; Transplantation; Transplants & implants; ultra‐structural characterization
• ISSN: 1059-910X; 1097-0029
• DOI: 10.1002/jemt.24447

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. hiPSC‐derived kidney organoids show remarkable morphological development of nephron segments upon transplantation. We performed ultrastructural characterization of these nephrons using high spatial resolution transmission electron microscopy (TEM), and observed formation of the glomerular filtration barrier, microvilli in the proximal tubule, and subsegmentation in the connecting tubule, and compared these findings to human kidney.