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This research employs ecosystem and strategic fit theories to understand incubators’ relationships with other ecosystem actors by investigating the elaboration of specialization, diversification, and co-opetition strategies. In the entrepreneurial support ecosystem, incubators act as intermediaries, bridging the gap between tenants and their external environment. These strategically led organizations offer a unique perspective for exploring the strategic fit differentiation engine through a holistic approach. A qualitative study reviews 48 semi-structured interviews derived from five case studies of the main incubator types. The results show that incubators differentiate themselves by employing various combinations of individual and collective strategies consisting of individual, organizational, and environmental factors to achieve ecosystem benefits. These findings provide a cross-level understanding of the entrepreneurial support ecosystem for all ecosystem actors and enable the implementation of appropriate generic strategies.