National culture and its various dimensions seem to affect various aspects of our lives. However, the role of national culture in the development of global literacy is still unclear. The aim of the paper is to evaluate whether the existence of cultural differences between Slovakia and Czechia can lead to differences in the level of global literacy of university students. The questionnaire-based survey was conducted on a sample of Slovak and Czech university students studying economics and business. The results indicate that despite significant cultural differences between the two nations in terms of some dimensions of Hofstede´s framework, no significant differences in the level of global literacy between both groups of students were detected. It seems that national culture does not play an important role in the development of global literacy and/or the cultural differences between Slovakia and Czechia are not so significant. These considerations deserve further investigation.