The Regional Centre of Expertise (RCE) Rhine-Meuse is one of the first seven pioneer RCEs and the first in Europe. It was established in 2004 after a preparation period that began in 2001. The period 2005–07, which is described and analysed in this article, was when the RCE established a position in the region and proved its added value. The RCE received funding from the European Regional Development Program to start up and from the Province of Dutch Limburg for 2006–07, enabling it to build a regional network with a central office, to set up an Internet-based communication and knowledge distribution system and to initiate its first pilot projects. Evaluation of the first two years of activities showed that serving the complex Euregion, Rhine-Meuse proved to be difficult in practice. This article focuses on stakeholder issues, the maturing of the RCE’s management system and its position in the region, and concludes with a perspective for the future.