Public universities as institutions of public sector have very significant position at the market of tertiary education in the Czech Republic. Universities and their activities had been negatively impressed with the communistic regime in the same way as activities of other institutions in the Czech Republic till 1989. First private universities appeared at the Czech market of tertiary education ten years later. Their activities at the market of tertiary education gave rise to first signs of competitive fight that intensified during next years. Public universities have begun to employ in their activities methods that were used primarily by private sector. One of methods that were originally intended only for private sector is mapping and analyzing of stakeholders. This article focuses on analyzing of stakeholders of public universities. The initial groups of stakeholders of public universities were defined on the basis of the literature search and approved by the authors’ survey. The aim of this survey was to define basic groups of stakeholders and their priorities according to the opinion of public universities.