Abstract: SA-PO0019
A Holistic Human Tissue-Derived Air-Liquid Interface Kidney Organoid System
Session Information
- Development, Stem Cells, and Regenerative Medicine
October 24, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 700 Development, Stem Cells, and Regenerative Medicine
Authors
- Gaeta, Xavier, Stanford University, Stanford, California, United States
- Sutha, Ken, Stanford University, Stanford, California, United States
- Guo, Alyssa J., Stanford University, Stanford, California, United States
- Kuo, Calvin J., Stanford University, Stanford, California, United States
Background
Our understanding of human kidney injury and diseases is limited by a lack of bona fide tractable human kidney culture systems in vitro.
Existing 2D systems and many iPSC organoids do not capture mature kidney physiology or cellular diversity. Adult tissue-derived organotypic cultures could better model injury and rejection by maintaining native architecture and immune–stromal crosstalk. This is especially relevant in the study of diseases including glomerulonephritis or transplant rejection. We aim to adapt air-liquid interface organoid methods to culture human kidney tissue with resident immune cells to study mechanisms of allograft rejection and support therapy development.
Methods
We adapted air–liquid interface culture methods to grow adult human kidney tissue–derived organoids from nephrectomy tissue and excess native or allograft biopsy material according to the Stanford IRB. Mechanically dissociated tissue was embedded in collagen I in transwell inserts and screened across defined growth factor conditions. We used a Wnt3a, Noggin, and Rspo-1 base supplemented with EGF for prolonged culture. Cystic outgrowths appeared by two weeks, and organoids were maintained long-term with periodic passage.
Results
Marker expression and single-cell RNA-seq show multilineage cells can be maintained in culture and continue to proliferate without nephron progenitor markers reappearing. Organoids can be passaged into fresh collagen and are amenable to freezing and thawing. They contain epithelium, vasculature, glomeruli, stroma, and immune cells. In ALI organoids derived from transplant biopsies, both donor epithelium and recipient immune cells are captured, and can be stimulated with anti-HLA antibodies or T-cell activating agents. These stimulated immune cells demonstrate higher motility in culture, induce epithelial cell death, and secrete inflammatory cytokines into the transplant milieu.
Conclusion
In conclusion, we developed and present here an adult human kidney tissue-derived air-liquid interface organoid system that preserves diverse, mature kidney and immune cell populations. These organoids expand long-term, can be passaged and cryopreserved, and hold the promise of enabling mechanistic studies of kidney injury, glomerulonephritis, and transplant rejection to enable deeper understanding of and therapeutic interventions for human kidney disease.
Funding
- Other NIH Support