This paper discusses the relationship between the entrepreneurial ecosystem and stakeholder groups involved in sustainable development of regions. Findings from empirical research of Latvian regions brought researchers towards improvement of the entrepreneurial ecosystem model. This paper develops an ecosystem model that assists in the planning and designing of regional sustainable development. Findings from empirical research might help us to identify additional contributions of stakeholder groups for sustainable development of regional entrepreneurial ecosystems. The paper is aimed at improving understanding of entrepreneurial ecosystem structure and suggesting ways for individual companies to exploit the components of entrepreneurial ecosystems. By summarizing the research results, the impact of stakeholder groups involved in the sustainable development of a regional entrepreneurial ecosystem is explained.