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Abstract: FR-PO916

Evaluating the Functional Response of a Barrier-Free Glomerulus-on-a-Chip System to Injury: A Novel Tool for Renal Personalized Medicine

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 501 Development, Stem Cells, and Regenerative Medicine: Basic

Authors

  • Marcheque, Julia, Children''s Hospital Los Angeles, Los Angeles, California, United States
  • Cravedi, Paolo, Mount Sinai, New York, New York, United States
  • De Filippo, Roger E., Childrens Hospital Los Angeles, Los Angeles, California, United States
  • Perin, Laura, Childrens Hospital Los Angeles, Los Angeles, California, United States
  • Da Sacco, Stefano, Children's Hospital Los Angeles, Los Angeles, California, United States
Background

With increasing rates of renal failure, there is an urgent need of implementing new strategies for understanding glomerular pathophysiology and for developing efficient patient-specific drug screening tools. To this regard, we recreated an in vitro 3D multicellular platform that allows fluid perfusion and proper interactions between podocytes and glomerular endothelial cells (hGEC). This glomerulus-on-a-chip system can be used to study the complex architecture and function of the glomerular filtration barrier (GFB).

Methods

Human amniotic fluid derived podocytes (hAKPC-P), human immortalized podocytes (iPod), primary human podocytes (pPod) and human fibroblasts (negative control) were seeded on microfluidic chips (OrganoplatesTM) with hGEC. Immunofluorescence was performed for podocyte, endothelial and GBM markers. Barrier selective-permeability was investigated. Feasibility of the system for high throughput screening and disease modeling was evaluated by measuring permselectivity following Puromycin Aminonucleoside (PAN) injury or culture with serum from FSGS (focal segmental glomerular sclerosis) and membranous nephropathy (MN) patients.

Results

We have developed an innovative, barrier-free, glomerulus-on-a-chip system. hAKPC-P, iPOD and pPOD formed a slit diaphragm-like structure expressing nephrin and podocin. CD31-expressing hGEC formed capillary-like structures. De-novo deposition of GBM components such as collagen IValpha3,4,5 and laminin alpha5 was confirmed for hAKPC-P and pPOD but not iPOD. Albumin permselectivity was confirmed on chips with hAKPC-P, pPOD and less efficiently for iPOD while use of fibroblast lead to marked albumin leakage. Permselectivity was impaired following PAN administration as confirmed by our experiments. In the presence of MN serum (but not from healthy controls or FSGS) IgG-mediated injury to the barrier caused albumin leakage, suggesting an MN specific mediated damage.

Conclusion

We have successfully developed a glomerulus-on-a-chip system that closely mimics the GFB and provides a powerful tool for studying renal regenerative, disease mechanisms and toxicity effects. In conclusion, this system will increase our ability to individualize treatments and drug susceptibility, thus ultimately benefiting patients affected by renal failure.

Funding

  • Private Foundation Support