The size of window openings is widely acknowledged as an important factor in our spatial perception. However, little is known about how the perception and preference of windows changes between countries, leaving a gap of knowledge regarding the applicability of research findings across latitudes. This article presents the outcomes of a study investigating regional differences in the perception of spaces with varying window size (small, medium, and large), space size (small and large), spatial context (working and social), and sky type (overcast and two types of clear sky). As the regional differences were the main studied factor, the study was performed in Norway, Switzerland, and Greece, representing northern, central, and southern European latitudes, respectively, and used virtual reality as a means to replicate the same experiment in different locations. In total, 406 participants evaluated eight spatial attributes using an 11-point Likert-type scale. Results indicated that regional differences could be observed in the participants’ responses, with significant differences in how pleasant and calm the space was perceived, found not only between participants in Greece and Norway in all the studied window sizes, but also between Greece and Switzerland for the medium and large windows, indicating that even small variations in latitude within Europe can affect the spatial perception. The findings of this study reveal that spaces with specific fenestration characteristics might not induce the same response across different latitudes in Europe, and thus, have important implications for daylighting and architectural design, which would motivate the use of region-specific parameters.