Purpose: The paper provides a theoretical foundation of entrepreneurial ecosystems with strong focus on start-up ecosystem. The empirical purpose of the study is to identify the key actors, institutions and organisations which can provide a nurturing environment and services for the creation of the start-up ecosystem of the city of Poznań with regard to business-science- government collaboration. Design/methodology/approach: The research applies the method of in-depth semi-structured direct interview with experts in the field of support services to potential founders and start-ups with academic origin, the case study method, participatory observation, and reflection. The explorative qualitative study uses both descriptive and explanatory techniques. Findings: The research findings provide insight in the real nature of the local start-up ecosystem presenting its key stakeholders, the scope of their support and ways of creating a conducive environment for potential founders and start-ups. The research results highlight the importance of studying the interdependencies among key actors in the start-up ecosystem in order to provide them with necessary resources and to stimulate the synergy effect. Research limitations/implications: Research limitations resulting from the analysis of a purposefully-selected case of the start-up ecosystem do not allow formulating general conclusions. Nevertheless, it illustrates a real business practice and challenges of the development of the specific entrepreneurial ecosystem. Future research line will concern an in-depth analysis of the most critical obstacles in the process of start-up creation as well as the assessment of the interdependencies among the key stakeholders of the start-up ecosystem to look for more effective cooperation. Practical implications: The synthesis of the current reflections on entrepreneurial ecosystems and the research findings reflected here can benefit both employees of business incubators, researchers, and entrepreneurship teachers and become an inspiration for further analysis and extended research on problems associated with increasing the efficiency and sustainability of local start-up ecosystems and the need to build valuable relationships with key stakeholders. Originality/value: The originality of the conducted exploratory research lies in presenting the real business practice and challenges of the development of the specific entrepreneurial ecosystem, and thus contribute to the discussion on the dilemmas associated with conducting the more effective practice-oriented research on start-up ecosystems.