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Here, we present a proof-of-concept co-culture platform to promote vascularization of kidney organoids by integrating stromal vascular fraction cells, collagen sponge scaffolds, porcine kidney decellularized extracellular matrix (dECM), and dynamic culture in a perfusion-based U-CUP bioreactor. We systematically evaluated the effects of serum supplementation, scaffold pre-vascularization, dECM encapsulation, and flow conditions on organoid–vascular integration. Low-level serum supplementation (1.5% FBS) sustained vascular networks without compromising organoid morphology. Perfusion supported vascular expansion; however, while a low flow rate comparable to interstitial tissue flow (0.01 mL/min) preserved organoid morphology and glomerular-like structures, a higher flow rate optimized for vascularization (0.47 mL/min) disrupted overall organoid architecture. Encapsulation within dECM preserved glomerular morphology and supported vascular infiltration, whereas co-culture with stromal vascular fraction cells enabled putative podocyte–endothelial interactions. Finally, exposure to the calcineurin inhibitor tacrolimus revealed dose-dependent cytotoxicity and characteristic nephron and vascular injury, demonstrating the model’s utility for nephrotoxicity screening. 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