Given the dynamics of the business environment, training in teamwork competence is becoming increasingly important vis-à-vis improving entrepreneurship and innovation development processes in a multidisciplinary context. In this paper, we estimate differences in differences (DID) regressions to analyse the effect of cooperative learning on teamwork competence for a sample of individuals whose training is geared towards acquiring key competences such as entrepreneurship and innovation. A quasi-experimental design is used with a treatment and a control group of two cohorts of individuals. Our results show a significant effect of cooperative learning on collective efficacy, planning, establishment of objectives, problem solving and conflict management. We contribute to student training in a competence that is highly valued in the professional sphere and to current understanding of what effect cooperative learning has from its different components in teamwork training.