Abstract: SA-OR14
Microfluidic Tissue Chip Technologies to Study Normal and Disease Podocyte Behaviors
Session Information
- Bioengineered Model Systems and Insights in Kidney Development and Function
October 26, 2024 | Location: Room 2, Convention Center
Abstract Time: 05:10 PM - 05:20 PM
Category: Bioengineering
- 400 Bioengineering
Authors
- Subramanian, Balaji Karthick, Beth Israel Deaconess Medical Center, Boston, Massachusetts, United States
- Pollak, Martin, Beth Israel Deaconess Medical Center, Boston, Massachusetts, United States
Background
Unraveling the complex behavior of healthy and diseased podocytes by analyzing the changes in their unique arrangement of foot processes, slit diaphragm, and three-dimensional (3D) morphology is a long-standing goal in kidney-glomerular research. The complexities surrounding podocyte accessibility in animal models and growing evidence of differences between human and animal systems have compelled researchers to look for alternate approaches to study podocyte behaviors. With the advent of bioengineered models, an increasingly powerful and diverse set of tools is available to develop novel podocyte culture systems.
Methods
Using multi-layer photolithography, we created a microfluidic chip to co-culture podocytes with endothelium. We used normal, INF2, and APOL1 FSGS patient-sourced induced pluripotent stem cells (IPSCs) to prepare podocytes and endothelial cells. We derived podocytes from kidney organoids while we directed the IPSCs differentiation to prepare endothelial cells. Unidirectional fluid flow for various time frames and shear stress were introduced through the system, and changes in the podocyte structure and function were examined. Changes in podocyte morphology, marker protein expression and localization, and transport properties were assessed and compared between normal and FSGS disease tissue chips.
Results
Endothelial cells formed a vessel in our chip system with proper marker protein (VE-cadherin) localization and barrier function. When podocytes were introduced to the chip system, they migrated toward endothelial cells and interacted with the outer curvature of endothelial vessels, thus recapitulating the invivo-like arrangement. Tissue characterization analysis of the tissue–-system confirmed fenestration-like structures in endothelium and foot processes-like extension in the podocyte. More importantly, podocytes recapitulated defective cell spreading, actin arrangement, and transport phenotype in the mutant-INF2 and APOL1 risk variant-based tissue chips.
Conclusion
We have developed and validated a model system to study podocyte behaviors ex vivo. Our platform will allow mechanistic studies and drug testing in an invivo-like environment. Establishing the tissue system using human iPSC-derived endothelial cells and podocytes enables researchers to study disease and test therapeutics in a personalized manner.
Funding
- NIDDK Support