Purpose:The empirical purpose of the study is to identify the key actors of the local universitybased start-up ecosystem and assess the forms of support (substantive, financial and organizational) they provide to create a conducive environment for academic start-ups. Design/Methodology/Approach: The paper draws on a broad management science literature review, covering various approaches to the concept of entrepreneurial ecosystems, using the following methods: exploration, interpretation, comparing, analysing critical factors, and inferring. The empirical 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 university-based start-up ecosystem presenting its key actors (stakeholders), the forms of their support for start-up incubation and ways of creating a conducive environment for potential founders and start-ups, as well as critical factors of entrepreneurial ecosystems. Practical Implications: The synthesis of the current reflections on entrepreneurial ecosystems can contribute to the discussion on the dilemmas associated with increasing the efficiency and sustainability of local start-up ecosystems and the need to build valuable relationships with external stakeholder. 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 this problem. Originality/Value: The originality of the conducted exploratory research lies in presenting the key actors, their interrelationships, and achievements of the specific university-based start-up ecosystem as well as the supporting activities and formats provided by the university business incubator to create a favourable environment for academic start-ups. The study highlights the mutual relationships within the start-up ecosystem and critical factors which are crucial to stimulate and support ambitious entrepreneurship in the region.