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Abstract: FR-PO357

Human Collecting Duct Tubuloids of Principal Cells Derived From Ureteric Bud Organoids as Differentiated Cell Models for Physiology and Disease Modelling

Session Information

Category: Development‚ Stem Cells‚ and Regenerative Medicine

  • 500 Development‚ Stem Cells‚ and Regenerative Medicine


  • Di Mise, Annarita, Universita degli Studi di Bari Aldo Moro, Bari, Puglia, Italy
  • McCracken, Kyle, Boston Children's Hospital, Boston, Massachusetts, United States
  • Patel, Ankit B., Vertex Pharmaceuticals Incorporated, Boston, Massachusetts, United States
  • Valenti, Giovanna, Universita degli Studi di Bari Aldo Moro, Bari, Puglia, Italy
  • Ichimura, Takaharu, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Bonventre, Joseph V., Division of Renal Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States

3D kidney cellular models, organoids or tubuloids that recapitulate tissue-specific pathophysiology, including disease phenotypes after genome editing, may represent useful tools for kidney disease modeling and drug screening. We report here the first model of tubuloids originated from 3D culturing of human collecting duct (hCD) cells obtained from pluripotent stem cells-derived ureteric bud organoids. These cells generate functional epithelia characterized by amiloride-sensitive transepithelial potential and high transepithelial resistance when evaluated in two-dimensional transwells. These functional properties of hCD principal cells are unique with no other comparable cell line available in the literature.


To generate hCD cell tubuloids, cells were cultured as monolayer (2D) or grown in Geltrex diluted in 2% serum DMEM/F12. After 1-2 days in Geltrex, hCD cells formed aggregates which developed into more complex 3D tubular structures, which detached from the original aggregates.


Both 2D cultures and 3D tubuloid cells expressed AVPR2 and AQP2 gene transcripts by qPCR. The water channel, AQP2, was poorly expressed in 2D cells as assessed by immunofluorescence with higher expression in tubuloids primarily in the intracellular vesicles. Activation of the V2R with dDAVP induced AQP2 relocation from the intracellular vesicles to the apical membrane of hCD-tubuloids, but not in 2D cells. Long-term exposure of hCD-tubuloids to dDAVP induced the growth of cysts. In addition, hCD tubuloids expressed PC2, a protein which is mutated in ~15% of individuals with Autosomal Dominant Polycystic Kidney Disease (ADPKD).


Since the collecting system plays an important role in regulating sodium, potassium, and acid-base homeostasis as well as disease states such as polycystic kidney disease and nephrogenic diabetes insipidus, the generation of hCD-tubuloids with functional properties of hCD principal cells represents a unique model for disease modelling in human cells in vitro.


  • NIDDK Support