Background and Aims: We have optimized the isolated perfused mouse kidney (IPMK) model for studying renal vascular and tubular function in vitro using 24-28 g C57BL6J mice; the wild type controls for many transgenic mice. Methods and Results: Buffer composition was optimized for bovine serum albumin concentration (BSA). The effect of adding erythrocytes on renal function and morphology was assessed. Autoregulation was investigated during stepped increases in perfusion pressure. Perfusion for 60 min at 90-110 mmHg with Krebs bicarbonate buffer containing 5.5% BSA, and amino acids produced functional parameters within the in vivo range. Erythrocytes increased renal vascular resistance (3.8 ± 0.2 vs 2.4 ± 0.1 mL/min.mmHg, P < 0.05), enhanced sodium reabsorption (FENa = 0.3 ± 0.08 vs 1.5 ± 0.7%, P < 0.05), produced equivalent glomerular filtration rates (GFR; 364 ± 38 vs 400 ± 9 μL/min per gkw) and reduced distal tubular cell injury in the inner stripe (5.8 ± 1.7 vs 23.7 ± 3.1%, P < 0.001) compared to cell free perfusion. The IPMK was responsive to vasoconstrictor (angiotensin II, EC50 100 pM) and vasodilator (methacholine, EC50 75 nM) mediators and showed partial autoregulation of perfusate flow under control conditions over 65-85 mmHg; autoregulatory index (ARI) of 0.66 ± 0.11. Angiotensin II (100 pM) extended this range (to 65-120 mmHg) and enhanced efficiency (ARI 0.21 ± 0.02, P < 0.05). Angiotensin II facilitation was antagonized by methacholine (ARI 0.76 ± 0.08) and papaverine (ARI 0.91 ± 0.13). Conclusion: The IPMK model is useful for studying renal physiology and pathophysiology without systemic neurohormonal influences.