This paper examines the diffusion of entrepreneurial activities among Italian universities, the evolution of the organizational models implemented to facilitate such activities and the commitment of the universities to the Third Mission of social and economic development. As previous analyses have shown, Italian universities have only recently moved towards the valorisation and exploitation of their scientific knowledge. In contrast to the varied and in some cases openly hostile attitudes of the past, there now seems to be greater acceptance of and a more positive approach towards entrepreneurship. Many universities have introduced innovations both in the organizational processes and activities of scientific research and in the management of research results. Because empirical evidence is very limited, the authors have used a longitudinal analysis to contribute to debate on the subject, with a discussion of the evolution of universities currently engaged in such activities and identification of possible changes in their cultures. (Contains 2 notes, 4 tables, and 1 figure.)