Over the last several decades the traditional role of universities in their unique mission in education and research changed to include also the so-called third mission consisting in their active involvement in the socio-economic development of the society. Publicly available data from annual economic report of Czech public universities were analysed to demonstrate the universities´ ability to raise funding from off-budgetary sources which belongs to main characteristics of entrepreneurial university. Off-budgetary sources, where revenues for internal services prevailed, made in 2016 around 16 % of the total income of Czech public universities. The off-budgetary sources include in particular charges for accommodation and board, fees for entrance exam, tuition fees for students that exceed the standard length of studies by more than one year, and tuition fees for studies in a foreign language. The last fees make up to three quarters of all offbudgetary sources at universities that offer studies at medical faculties. The universities´ ability to execute the third role was due to limited availability of data accessed only partially by analysing revenues from knowledge transfer in supplementary activities that involve contractual research, income from licences, educational courses on demand, consultations and counselling. Contractual research in supplementary activities was the most important among the listed activities making up to 8 % of all off-budgetary sources revenues. In international comparison the Czech public universities raise about one half of revenues for R&D from entrepreneurial sources than EU-28 average and only one third compared to German universities. Revenues from license agreements are low making around 0.1 % of all revenues for R&D.