The sustainable mobility sector has been recently identified by the Ministry of Economy as one of the most promising directions in Luxembourg’s strategy for the diversification and greening of the economy. The eCoBus project is the first research activity linked to the initiative of the Ministries of Economy and Infrastructure, Volvo Buses, and Sales-Lentz to create in Luxembourg a test arena for sustainable public transport. This initiative inaugurates in the second half of 2016 with a pilot deployment of a plug-in hybrid electric bus system with on-route charging. With eCoBus the University of Luxembourg (UL) teamed up with the Luxembourg Institute of Science and Technology (LIST) to design and assess a system approach exploiting the potentials of the new Cooperative Intelligent Transportation Systems (C-ITS) paradigm to meet the requirements of the next generation Public Transport (PT) systems. Such systems put an emphasis on green vehicles, increased comfort, and operating costs reduction. However, they introduce additional complexity—for instance electric buses need to periodically recharge their batteries on-route using dedicated infrastructure. This not only can impact service level, but also extend operating costs with complex electric charges. In this project we argue that the aforementioned requirements—unsupported by current control methods—can be met by introducing new strategies explicitly optimising the interactions within the PT ecosystem components consisting of PT vehicles, traffic signals, and electric bus charging infrastructure. The main novelty of eCoBus is that these strategies combine cooperation and negotiation within the whole PT ecosystem, enabled by C-ITS connectivity. To achieve these goals, we mainly rely on vehicle control rather than on the use of prioritised treatment at traffic signals, which in busy urban scenarios can have negative impacts on overall traffic. The main research challenges are in formulating and solving complex multi-objective optimisation problems involving game-theoretical aspects. The proposed system will be tested and evaluated not only in extensive simulations but also in real-world controlled experiments supported by our PT industry partners—Volvo Buses and Sales-Lentz (PT operator).