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In the context of the global economy and growing competition within the country and on the international market, the development of entrepreneurial ecosystems is an important factor for sustainable economic growth. The author distinguishes between the concepts of “entrepreneurial ecosystem” and “cluster”. In a new view of entrepreneurial ecosystems as dynamic structures that go through the stages of birth, formation, development, maturity, stabilization, renewal or decline, according to the author’s methodology, special attention is paid to the mechanism of prolongation of growth stages due to the influence of clusters. The integration of life cycle theory and clusters provides a new perspective on entrepreneurial ecosystems through the lens of clusters. The novelty lies in the role of clusters as an effective approach that promotes interaction between business, government and science to accelerate innovation processes, create sustainable cooperation networks, attract investment and reduce barriers to entry of new entrants to the market. To substantiate the decision to select the largest entrepreneurial ecosystems, a comparative analysis was carried out for the Novosibirsk Region in the context of the regions of the Siberian Federal District, for the Republic of Tatarstan – in the context of the regions of the Volga Federal District. An analysis of the economic sectors of the studied regions revealed that entrepreneurial companies not only operate in the main ones, but also form related industries. It is revealed that the industry association leads to the creation of new industries of the sixth technological order and the production of competitive goods and services, which creates an additional need for investments for the development of innovation activities by entrepreneurial ecosystem companies. The use of the OTSW analysis tool (according to V. L. Quint’s method) is substantiated. This article will help determine the approach when making a management decision to increase the vital activity of the entrepreneurial ecosystem. The formation of clusters will improve the interaction between ecosystem participants, develop entrepreneurship and stimulate innovation in the country’s regions.