A hybrid off-lattice agent-based model was developed to reconstruct the tumor oxygenation landscape based on histology images and simulated interactions between vasculature and cells with microenvironment metabolites. Here, we performed a robustness sensitivity analysis of the model’s physical and computational parameters. We found that changes in the computational domain boundary conditions, the model initial conditions, and the Michaelis constant were negligible and, thus, did not affect the model outputs. The model was also not sensitive to small perturbations of the vascular influx or the maximum consumption rate of oxygen. However, the model was sensitive to large perturbations of these parameters and changes in the tissue boundary condition, emphasizing an imperative aim to measure these parameters experimentally.