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Description
Introduction: The glomerular filtration barrier (GFB) is a highly specialized structure responsible for blood filtration in the kidney; is composed of podocytes, glomerular basement membrane and fenestrated endothelial cells. Dysfunction of this barrier is a hallmark of many glomerular diseases. However, the current systems in vitro cannot mimic efficiently the 3D organization and the biochemical and mechanical properties of human GFB. This study aimed to develop a biomimetic human GFB in vitro for studying glomerular physiology and pathology.
Methods: To engineer the GFB, 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 of the transwell integrated into a 3D millifluidic chip. To more faithfully mimic the physiological condition, the polyester membrane was coated with natural glomerular basement membrane components (i.e. different collagen types, fibronectin etc.) or completely replaced by an extracellular matrix gel. This system was linked to a peristaltic pump to mimic physiological shear stress on cells and to promote cell adhesion, alignment, and maturation.
Results: Our results demonstrated the successful establishment of a 3D biodevice comprising hiPSC-derived podocytes and glomerular endothelial cells separated by a self-assembled basement membrane. Characterization by qPCR and immunofluorescence confirmed the expression of key maturation markers for the GFB. Functional integrity of this barrier was validated through selective permeability assays. To model disease, we induced nephrotoxicity using Adriamycin and incorporated iPSCs 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.
Discussion: This GFB platform represents a significant step forward in the development of human-relevant models for renal research in vitro. It provides a versatile tool for investigating disease mechanisms, enabling personalized medicine approaches, and advancing drug discovery.
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