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Abstract: FR-OR018

Human Induced Pluripotent Stem Cell (iPSC)-Derived Glomerulus-on-a-Chip for Disease Modeling and Drug Testing

Session Information

Category: Bioengineering

  • 400 Bioengineering

Authors

  • Locatelli, Laura, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
  • Fiorentini, Eleonora, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
  • Petracca, Benedetta, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
  • Longaretti, Lorena, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
  • Lavecchia, Angelo Michele, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
  • Remuzzi, Giuseppe, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
  • Xinaris, Christodoulos, IRCCS Istituto di Ricerche Farmacologiche Mario Negri Centro Anna Maria Astori, Bergamo, Lombardy, Italy
Background

The glomerular filtration barrier (GFB) - comprising podocytes, the glomerular basement membrane (GBM), and fenestrated endothelial cells - is essential for selective blood filtration in the kidney. Dysfunction of this barrier is a hallmark of many glomerular diseases. However, human in vitro models that faithfully replicate its complex architecture and function remain scarce.

Methods

To engineer a glomerulus-on-a-chip, human podocyte and glomerular endothelial cell lines were first used to optimize experimental conditions and ensure appropriate cell adhesion and viability. hiPSC-derived podocytes and glomerular endothelial cells were then co-cultured on opposite sides of a porous membrane, coated with natural GBM components, within a 3D millifluidic device. Physiological shear stress was applied via a peristaltic pump to promote cell adhesion, alignment, and maturation.

Results

Our results demonstrated the successful establishment of a two-layer cellular structure—comprising iPSC-derived podocytes and glomerular endothelial cells—within a millifluidic device, along with the deposition of glomerular basement membrane (GBM) components. Characterization by qPCR and immunofluorescence confirmed the expression of key maturation markers for podocytes, endothelial cells and GBM. Functional integrity of the glomerular filtration barrier (GFB) was validated through selective permeability assays. To model disease, we induced nephrotoxicity using Adriamycin and incorporated hiPSCs derived from patients with Alport syndrome. Perm-selectivity assays revealed increased permeability in both disease conditions compared to healthy controls. Analysis of samples from Adriamycin-treated and AS-derived models further identified disease-specific alterations in GFB structure and function.

Conclusion

This hiPSC-based glomerulus-on-a-chip provides a robust and physiologically relevant human model for studying glomerular biology and pathology. It enables patient-specific disease modeling, nephrotoxicity testing, and evaluation of therapeutic efficacy, supporting both mechanistic studies and drug development pipelines.

Digital Object Identifier (DOI)