Various techniques of stentless aortic valve implantation with or without wall components exist. We investigated the in-vitro performance of stentless valves without or with aortic wall removal mimicking root versus subcoronary implantation. Glutaraldehyde-preserved stentless aortic valves (gpSVG), cryo-preserved human homografts (cpHG), cryo-preserved xenografts (cpXG), and fresh xenografts (fXG) were used. Valves were mounted as full roots or trimmed in a mock circuit. Mean transvalvular gradient (MTVG, mmHg) was measured. Distensibility was quantified using post-systolic backflow volume (BV, ml) - after valve closure. Function was visualized by means of a high-speed camera. Glutaraldehyde-preserved valves exhibited higher MTVG than cryo-preserved or fresh substitutes. After trimming, cpHG, cpXG, and fXG demonstrated marked reduction of MTVG (cpHG: 7.6-5.2 mmHg; cpXG: 6.7-4.9 mmHg; fXG: 8.4-5.2 mmHg). In contrast, after trimming gpSVG exhibited a significant increase of MTVG (7.1-9.2 mmHg). BV remained constant. Visualization indicated maintained distension of all valves and types of all sizes after trimming. In fresh and cryo-preserved grafts, aortic wall trimming resulted in significantly improved systolic performance while glutaraldehyde-preserved stentless valves demonstrated systolic impairment after wall resection. Subcoronary implantation of fresh or cryo-preserved aortic valves may therefore be preferred. In contrast, glutaraldehyde-preserved valves are dependent on wall suspension and may therefore be implanted as a root.