Abstract: SA-OR12
Engineering Scalable Vascularized Kidney Organoids with the Integration of Human Endothelial Cells for In Vivo Glomerular Filtration
Session Information
- Bioengineered Model Systems and Insights in Kidney Development and Function
October 26, 2024 | Location: Room 2, Convention Center
Abstract Time: 04:50 PM - 05:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 600 Development, Stem Cells, and Regenerative Medicine
Authors
- Tekguc, Murat, Massachusetts General Hospital, Boston, Massachusetts, United States
- Morizane, Ryuji, Massachusetts General Hospital, Boston, Massachusetts, United States
Group or Team Name
- Morizane Lab.
Background
Kidney organoids derived from human pluripotent stem cells have emerged as promising models for studying kidney disease pathogenesis and the development of novel therapeutics. However, previous conventional production methods rely on traditional static cell cultures, thereby limiting the scalability of organoids in the absence of vascularized glomeruli, a pivotal structure responsible for the kidney’s blood filtration function. Here, we present a practical dynamic-flow culture protocol using a stirred tank bioreactor (STR) with a delta-wing-shaped impeller, which can overcome the limitations of current methods by boosting large-scale manufacture of kidney organoids exhibiting glomerular vascularization.
Methods
1-Differentiation of human iPSCs (BJFF.6) and ESCs (H9) in both static and STR cultures
2-Treatment of STR organoids with the α2β1 integrin inhibitor
3-Generation and implantation of nephron sheets into immunocompromised NOD/SCID/IL-2 receptor common gamma chain-deficient (NSG) mice
4-Immunostaining, confocal microscopy, and 3D image analysis
5-Multiphoton Intravital Microscopy
6-Transmission Electron Microscopy (TEM)
7-qRT-PCR, RNA-seq Library preparation, sequencing, and RNA-seq Data Analysis
Results
The incorporation of a dynamic culture environment in delta-wing stirred bioreactors has significantly enhanced the glomerular vascularization and maturation of kidney organoids via mechanosensory integrin α2β1. This robust, scalable, labor- and cost-efficient platform provided large quantities of vascularized kidney organoids, enabling the fabrication of a nephron sheet with nephron numbers equivalent to those found in two rat kidneys. Intravital imaging of a nephron sheet implanted in a dorsal skinfold chamber of mice revealed in vivo filtration function with size selectability in the organoid glomeruli vascularized with human endothelia. In summary, we have developed a cost-effective mass-production method to manufacture functional vascularized kidney organoids, which has improved production efficiency by more than 50 times compared to conventional culture systems.
Conclusion
This work may represent a significant step towards bridging the gap between basic research and therapeutic applications for patients, paving the way towards facilitating the biofabrication of 3D large kidney tissues for kidney replacement therapy.
Funding
- Other NIH Support