Kidney Week

Abstract: TH-PO0042

Combinatorial Effects of Matrix Composition, Mechanical Stiffness, and Fluid Shear Stress on Induced Pluripotent Stem Cell-Derived Podocyte Maturation

Session Information

• Bioengineering: MPS, Flow, and Delivery
  November 06, 2025 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Bioengineering

• 400 Bioengineering

Authors

• Zarouk, Alexander A., Cleveland Clinic Lerner Research Institute, Cleveland, Ohio, United States

Alexander A. Zarouk, MS

• Beamish, Jeffrey A., University of Michigan Medical School, Ann Arbor, Michigan, United States

Jeffrey A. Beamish, MD, PhD

• Tran, Uyen, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio, United States

Uyen Tran

• Wessely, Oliver, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio, United States

Oliver Wessely, PhD

Background

Regenerative medicine efforts have focused to recapitulate the glomerular filtration barrier by differentiating induced pluripotent stem cells (iPSCs) into functional podocytes. Current differentiation protocols inadequately replicate the native glomerular mechanical and extracellular matrix (ECM) signals, likely limiting functional maturation. We hypothesize that combining defined ECM substrates with biomechanical stimuli will enhance podocyte maturation and stimulate ECM remodeling towards a mature glomerular basement membrane (GBM).

Methods

iPSC-derived nephron progenitor cells (NPCs) were seeded onto plates coated with GBM proteins/mimetics (Collagen I, Collagen IV, Laminin-521, Silk Sericin, Poloxamer 407) individually or in combination using either 3D hydrogels of tunable stiffness or 2D substrates (~7 kPa). Podocytes were subsequently differentiated under static conditions or orbital shear stress (50–200 rpm; 0.1–1.10 dyn/cm²). Expression of podocyte differentiation markers were quantified via qRT-PCR and validated by immunofluorescence staining.

Results

NPCs cultured on mechanically compliant matrices under low-magnitude shear stress showed increased expression of podocyte-specific markers (WT1, NPHS1, NPHS2) and notably induced collagen IV isoform switching towards COL4A3and COL4A5, indicative of GBM maturation. Podocyte differentiation marker gene expression was enhanced upon exposure to ECM substrates. ECM substrates mimicking native GBM components, specifically Collagen IV, enhanced podocyte differentiation compared to static or non-native ECM conditions. This was further improved by exposure to shear stress.

Conclusion

The integration of physiologically relevant ECM substrates and biomechanical stimuli significantly improves iPSC-derived podocyte maturation and ECM remodeling. These results highlight the necessity of accurately replicating glomerular microenvironmental conditions to facilitate effective podocyte differentiation. Applying multi-dimensional optimization strategies, such as Design-of-Experiments platforms, will be critical to refine protocols for generating functionally mature podocytes, supporting regenerative medicine applications and disease modeling.

Funding

• NIDDK Support

Digital Object Identifier (DOI)

• doi: 10.1681/ASN.2025sz0b629w