The modern university is undergoing a fundamental transformation, transitioning from a model consisting of monodisciplinary faculties and research-based education to a third-generation university model. A third-generation university is characterized by the presence of a third mission in addition to the traditional two missions associated with the transfer of information and the generation of new knowledge. The authors analyzed the works of Russian and foreign researchers devoted to the third mission of universities. Currently, there is no single concept describing this concept. The conducted theoretical analysis of sources allowed us to identify a set of characteristics of the third mission of the university associated with interdisciplinarity, international activities, development of channels of interaction with the external environment, the presence of an entrepreneurial ecosystem. The study of the practical experience of universities in the Samara region in terms of implementing the third mission proves the presence of a significant number of characteristics of the third-generation university, but does not allow us to conclude about a large-scale transition to the stage of the third-generation university. In general, the activities of universities are carried out in accordance with the Humboldt University model, but federal programs and projects lead higher education organizations to the need for further development in terms of a more systematic and in-depth implementation of the third mission. Further stages of the transition to a third-generation university may be associated with the modernization of the university structure with the aim of a larger-scale implementation of approaches characteristic of higher education at the next stage of evolutionary development.