Single-Pass Perfusion of the Mouse Kidney In Situ for Functional and Pharmacological Studies

​Whole-organ models that preserve integrated renal architecture while permitting controlled experimental manipulation are important tools for investigating renal physiology. This protocol describes perfusion of the mouse kidney while it remains anatomically in situ but is functionally isolated from the systemic circulation, allowing assessment of integrated renal function and acute pharmacological responses. Following anesthesia, the kidney is perfused in situ with Krebs-Henseleit bicarbonate buffer containing 1% bovine serum albumin using a normothermic, oxygenated, single-pass, constant-flow system. Venous effluent and urine are collected for assessment of renal function. Preparation performance was assessed using arterial inflow pressure, paired inflow and outflow measurements of pH, partial pressure of carbon dioxide (pCO₂), and partial pressure of oxygen (pO₂), and cortical vascular histology. These parameters remained stable during a 90 min experimental period. Infusion of 1 mM phloridzin, a non-selective inhibitor of sodium-glucose cotransporters, increased urinary glucose excretion compared with vehicle treatment. The protocol provides a stable kidney preparation with preserved renal filtration, excretory function, and metabolism for at least 90 min. As such, the model enables controlled delivery of test compounds and high-temporal-resolution sampling of perfusate and urine, providing a foundation for acute studies of integrated renal physiology and pharmacology.