The aim of the research in this chapter is to deepen our knowledge about the relationships between actors in a regional ecosystem through the lens of a business incubator – specifically, the ways through which the incubator interacts to foster the development of techno-entrepreneurship at the regional level. Given the explorative nature of this work, the study adopts a qualitative single case study method focusing on The Net Value incubator, in order to investigate its role in facilitating the development of techno-entrepreneurship within the Sardinian regional ecosystem (Italy). This research develops a theoretical framework showing the interactions between the actors of a regional ecosystem from the perspective of a business incubator. Furthermore, we advance five propositions to explain how the business incubator facilitates these interactions to foster techno-entrepreneurship. For practitioners and new techno-entrepreneurs, this study proposes an interesting framework which might help them to understand the strategic advantages arising from the relationships between multiple actors in a regional ecosystem, and, also, how these relationships can foster the development of techno-entrepreneurship within such an ecosystem.