Diverse aspects of de-agrarianization, which is manifested by the cessation or significant reduction in agricultural activities, have been clearly visible at the outskirts of large cities in Central Europe in recent decades. The key drivers behind this process include increased pressures to cover peri-urban agricultural land by new developments, inadequate protection of agricultural land, ineffective implementation of urban planning policies, low recognition of the importance of agriculture, and overall changes in people’s dietary habits. Urbanization pressures undoubtedly belong to the factors intensifying overall de-agrarianization, as urban farmers are usually not able to compete with other urban functions. This article focuses on more in-depth understanding of the driving forces behind de-agrarianization processes that are specific to post-socialist cities. As a case study, Brno, a second-tier city in the Czech Republic, was selected. In the first part, the conceptual framework and drivers of de-agrarianization are discussed specifically for the case of large Central European post-socialist cities. In the next part, we explore by means of a set of qualitative interviews the case study of the regeneration of the area of a former Cistercian monastery in Brno that was traditionally used for agricultural purposes, but recently was redeveloped for a university campus. Our findings signal procedural issues connected to the preservation of architectural heritage during the regeneration that frequently end up with only fragments being preserved. We also demonstrate a decline in the use of urban agricultural properties that are hastily transformed into a new urban environment under extremely strong urbanization pressures. We argue that even in economically prosperous cities with highly neoliberal competition between possible urban land uses, agriculture must be considered a relevant and highly important urban function and more protected by planning tools.