In this research, the results of the life cycle assessment of polyurethane (PUR) foams with different recycled polyol contents are presented. A methodological framework implementing laboratory activities directly into the life cycle assessment has been developed. Laboratory activities made the primary data related to the recycled polyol production available through the glycolysis of polyurethane scraps and the subsequent production and characterization of the foams. Five different formulations were analyzed with glycolyzed polyol content ranging from 0 to 100%. A comprehensive set of impact categories was considered. To ensure the robustness of the results, the influence of two different end-of-life allocation approaches was investigated, and the model was subjected to sensitivity and uncertainty analyses. Formulations with recycled content of 50 and 75% scored better environmental impacts compared to others. The main contributions to the overall impact resulted to be related to the production of isocyanate and virgin polyol. Physical characteristics such as density and thermal conductivity emerged as the main variables to be considered to minimize the overall environmental impacts of PUR foams.