Endothelial-Fibroblast Organoids Generated in Hydrogel Microchamber Arrays: A Translational Model of Vascular Dysfunction in Cardio-Kidney-Metabolic Disease
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- Cell / in-vitro
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- Not yet reviewed by a person · added Oct 4, 2026
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Abstract
[UNLABELLED] Cardio-kidney-metabolic disease comprises interconnected cardiovascular, renal, and metabolic disorders in which endothelial dysfunction drives microvascular and macrovascular injury. Although novel treatments improve clinical outcomes, the cellular and molecular mechanisms underlying these effects remain incompletely understood. We developed and characterized a three-dimensional (3D) endothelial-fibroblast co-culture platform to study endothelial morphogenesis and responses to metabolic and pharmacological stimuli. [METHODS] A hydrogel microchamber array (HMC) platform was used to generate reproducible endothelial spheroids and extracellular matrix embedded organoids. Endothelial cells were cultured alone or with fibroblasts. Morphometric changes were quantified over time following glucose exposure and treatment with the GLP-1 receptor agonist/insulin combination (Xultophy). [RESULTS] The HMC platform enabled reproducible formation of spheroids and organoids. Fibroblast co-culture enhanced endothelial organization and structural complexity. Glucose induced organoid expansion and reduced circularity, consistent with structural remodeling. Xultophy attenuated these changes and preserved organoid morphology. Responses were dependent on the sequence of stimulation, with partial mitigation of glucose-induced effects by Xultophy. Organoids demonstrated greater sensitivity than spheroids, highlighting the importance of extracellular matrix context. [CONCLUSIONS] The HMC platform provides a reproducible and physiologically relevant 3D system for investigating endothelial-stromal interactions, metabolic stress, and therapeutic modulation in cardio-kidney-metabolic disease.