Evaluation and simulation of the multiaxial mechanical behavior of native and engineered soft tissues is becoming more prevalent. In spite of this growing use, testing methods have not been standardized and methodologies vary widely. The strong influence of boundary conditions were recently underscored by Waldman [2002, J. Materials Science: Materials in Medicine13, pp. 933–938] wherein substantially different experimental results were obtained using different sample gripping methods on the same specimens. As it is not possible to experimentally evaluate the effects of different biaxial test boundary conditions on specimen internal stress distributions, we conducted numerical simulations to explore these effects. A nonlinear Fung-elastic constitutive model (Sun, 2003, JBME125, pp. 372–380, which fully incorporated the effects of in-plane shear, was used to simulate soft tissue mechanical behavior. Effects of boundary conditions, including varying the number of suture attachments, different gripping methods, specimen shapes, and material axes orientations were examined. Results demonstrated strong boundary effects with the clamped methods, while suture attachment methods demonstrated minimal boundary effects. Suture-based methods appeared to be best suited for biaxial mechanical tests of biological materials. Moreover, the simulations demonstrated that Saint-Venant’s effects depended significantly on the material axes orientation. While not exhaustive, these comprehensive simulations provide experimentalists with additional insight into the stress–strain fields associated with different biaxial testing boundary conditions, and may be used as a rational basis for the design of biaxial testing experiments.