In this exploratory study with a qualitative approach, the process and strategies for the formation of entrepreneurship and innovation ecosystems were analyzed in territories of low demographic density, starting from the projects: Vale do Tejo Technology Park, Living Lab Beira da Cova and Habitat of Business Innovation in the Strategic Sectors, respectively, in Abrantes, Fundão and Penela (Portugal). Interviews were carried out with nine social actors linked to the projects and the content analysis technique was used with support from the NVivo software. Based on empirical analysis and literature review, a conceptual framework was developed for the formation of entrepreneurship and innovation ecosystems, where the following strategies were outlined: (i) urban rehabilitation programs, (ii) fostering the culture of hospitality; (iii) increased use of endogenous resources, (iv) quality of life, (v) qualification of human resources and (vi) support for entrepreneurship and innovation. It is noteworthy that the proposed framework can contribute to the guidance of public policies and the development of entrepreneurship and innovation ecosystems in low-density territories.