Following the concept of ecosystem services, we propose in this article an interdisciplinary and participatory methodological framework for ecosystem services assessment and participatory decision-making in Mediterranean cultural landscapes linked with transhumant pastoralism. It is based on four sequential phases: 1. characterisation of the social-ecological network associated with transhumance, 2. preliminary identification and characterisation of ecosystem services, 3. evaluation of ecosystem services (in biophysical, socio-cultural, and economic terms), and 4. future scenario planning for the analysis of social conflicts related to ecosystem services use and trade-offs as well as the proposal of management strategies. Applying the framework to a case study on one of the major transhumance landscapes in Spain, we could identify and evaluate more than 30 ecosystem services. The framework facilitated the design of robust policy measures that aim to maintain this livestock raising model and its associated flow of ecosystem services. It also contributes to provide the basis for the implementation of adaptive co-management strategies.