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Kidney Week

Abstract: PO0667

An Accelerated Method of Podocyte Differentiation from Human Induced Pluripotent Stem Cells for Modeling Diabetic Nephropathy

Session Information

Category: Diabetic Kidney Disease

  • 601 Diabetic Kidney Disease: Basic


  • Bejoy, Julie, Vanderbilt University Medical Center, Nashville, Tennessee, United States
  • Woodard, Lauren Elizabeth, Vanderbilt University Medical Center, Nashville, Tennessee, United States

Podocytes are highly specialized visceral epithelial cells that maintain glomerular barrier function and play important roles during both kidney development and progression of glomerular disease. Mature podocytes extracted from mammalian kidneys are difficult to culture long-term, hindering research on podocytopathies. Establishing an alternative, inexhaustible source of podocytes would be a valuable tool for understanding the molecular mechanisms underlying specific podocytopathies and developing targeted therapies. The discovery of induced pluripotent stem cells (hiPSCs) led to several protocols for deriving podocytes from hiPSCs, which could potentially serve as an unlimited source of podocytes.


All the existing effective methods for hiPSC- derived podocytes either require lengthy culture times (~ 30 days) or expensive media. Therefore, we sought to develop a faster and less expensive method. We found a simple and effective method to derive podocytes from hiPSCs in twelve days of culture and at lower cost. Our method followed a stepwise protocol in which the hiPSC were differentiated into primitive streak followed by intermediate mesoderm using activation of Activin A and Wnt signaling. Intermediate mesoderm cells were treated with FGF9 to generate nephron progenitors, followed by a cocktail of established growth factors to finally derive mature podocytes.


The developed podocytes expressed podocyte markers including PODX, synaptopodin, MAFB, Nephrin at protein levels comparable to the existing methods. Flow cytometry analysis revealed that our method results in generation of ~80% mature glomerular podocytes. We confirmed the functionality of the hiPSC-derived podocytes via permeability assay for FITC-albumin uptake. Next, we treated the cells with media containing high glucose (100mM) to generate a hiPSC-derived podocyte model of diabetic nephropathy. The podocytes showed actin rearrangement upon treatment with high glucose, suggesting the ability of these cells to effectively model podocytopathies. In addition, treatment with high glucose resulted in increased cytotoxicity and reduced viability in the podocytes.


Altogether, we have discovered a faster and less expensive method of podocyte differentiation from hiPSCs, as well as a new tissue culture model of diabetic nephropathy for disease modeling.


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