The emergence of entrepreneurial university is the result of inner development of university, impact of external factors on university, and increasing importance of knowledge and innovation in society. This study aimed Identifying and ranking the Barriers to Entrepreneurial University Ecosystem Development in Iran. This was a based on the sequential exploratory mixed method and in two sections of qualitative and quantitative. In qualitative section used grounded theory method and in quantitative approach researchers used correlational method. The population consisted of and professors, managers of entrepreneurship centers, growth and science and technology parks, industry department authorities, and PhD candidates in Tehran, Beheshti and Allameh Tabatabei University. Using purposeful sampling method. Using purposeful sampling method, 20 people were selected as key informants. The data were collected through and semi-structured interview. two methods including reviewing accountable validity and external were used to obtain validity and reliability of data. The data were analyzed by open, axial, and selective coding. At initial stage, 96 conceptual propositions were obtained; in second stage, they were classified in 26 categories. At axial coding stage, 6 key items were counted. Financial, structural, research, administrative, communication, traditional education and organizational culture were approved using factor loading, extraction mean variance and high convergence validity (0. 4), z high coefficients (1. 96), coefficient of determination (R2) and goodness of fit (GoF= 0. 56).