Differentiated mouse kidney tubuloids as a novel in vitro model to study collecting duct physiology
Olde Hanhof, C. J.A.; Dilmen, E.; Yousef Yengej, F. A.; Latta, F.; Ammerlaan, C. M.E.; Schreurs, J.; Hooijmaijers, L.; Jansen, J.; Rookmaaker, M. B.; Orhon, I.; Verhaar, M. C.; Hoenderop, J. G.
(2023) Frontiers in Cell and Developmental Biology, volume 11
(Article)
Abstract
Kidney tubuloids are cell models that are derived from human or mouse renal epithelial cells and show high similarities with their in vivo counterparts. Tubuloids grow polarized in 3D, allow for long-term expansion, and represent multiple segments of the nephron, as shown by their gene expression pattern. In addition, human
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tubuloids form tight, functional barriers and have been succesfully used for drug testing. Our knowledge of mouse tubuloids, on the other hand, is only minimal. In this study, we further characterized mouse tubuloids and differentiated them towards the collecting duct, which led to a significant upregulation of collecting duct-specific mRNAs of genes and protein expression, including the water channel AQP2 and the sodium channel ENaC. Differentiation resulted in polarized expression of collecting duct water channels AQP2 and AQP3. Also, a physiological response to desmopressin and forskolin stimulation by translocation of AQP2 to the apical membrane was demonstrated. Furthermore, amiloride-sensitive ENaC-mediated sodium uptake was shown in differentiated tubuloids using radioactive tracer sodium. This study demonstrates that mouse tubuloids can be differentiated towards the collecting duct and exhibit collecting duct-specific function. This illustrates the potential use of mouse kidney tubuloids as novel in vitro models to study (patho)physiology of kidney diseases.
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Keywords: cell physiology, collecting duct, epithelial sodium transport, organoid, tubuloid, tubulopathy, Developmental Biology, Cell Biology
ISSN: 2296-634X
Publisher: Frontiers Media S. A.
Note: Funding Information: The authors acknowledge the support of the Dutch Kidney Foundation (17PhD04 and 19OK005), the partners of “Regenerative Medicine Crossing Borders” (RegMed XB), Powered by Health∼Holland, Top Sector Life Sciences and Health and the Gravitation Program “Materials Driven Regeneration,” funded by the Netherlands Organization for Scientific Research (024.003.013) and the Netherlands Organization for Scientific Research (NWO Veni Grant No: 091 501 61 81 01 36). The authors thank Prof. Koenderink for providing the Na+/K+-ATPase antibody and C. Bos (Department of Physiology, Radboud University Medical Center, Nijmegen, the Netherlands) for preparing the immunohistochemistry samples. The authors also would like to thank the Radboudumc Technology Center microscopy for their support. Finally, the isotope used in this research was supplied by the U.S. Department of Energy Isotope Program, managed by the Office of Isotope R&D and Production. Funding Information: The authors acknowledge the support of the Dutch Kidney Foundation (17PhD04 and 19OK005), the partners of “Regenerative Medicine Crossing Borders” (RegMed XB), Powered by Health∼Holland, Top Sector Life Sciences and Health and the Gravitation Program “Materials Driven Regeneration,” funded by the Netherlands Organization for Scientific Research (024.003.013) and the Netherlands Organization for Scientific Research (NWO Veni Grant No: 091 501 61 81 01 36). The authors thank Prof. Koenderink for providing the Na/K-ATPase antibody and C. Bos (Department of Physiology, Radboud University Medical Center, Nijmegen, the Netherlands) for preparing the immunohistochemistry samples. The authors also would like to thank the Radboudumc Technology Center microscopy for their support. Finally, the isotope used in this research was supplied by the U.S. Department of Energy Isotope Program, managed by the Office of Isotope R&D and Production. + + Publisher Copyright: Copyright © 2023 Olde Hanhof, Dilmen, Yousef Yengej, Latta, Ammerlaan, Schreurs, Hooijmaijers, Jansen, Rookmaaker, Orhon, Verhaar and Hoenderop.
(Peer reviewed)
