BACKGROUND Personal values influence nursing students' development of professional values, which affect professional outcomes, and how nursing students react to different situations. Personal values can be shaped by different factors, including culture, gender, and age. OBJECTIVE To explore personal values held by nursing students, and to verify if and how gender and year of study affect nursing students' personal values. METHODS A multicenter, cross-sectional study was used. METHODS The whole population of nursing undergraduate students available at the time was recruited from eight centers of two Universities, composing a sample of 947 students. Demographic data were collected and it was administered the Portrait Values Questionnaire. METHODS Ethical approval was obtained from the Institutional Review Boards of the University of the participating centers. RESULTS The study sample was mainly composed of young (92.6%, n = 877), female (77.3%, n = 732), Italian (95.8%, n = 907), and unmarried (98.6%, n = 934) nursing students. The most important value for nursing students, consistently through the years of nursing school, was Self-transcendence, which has the motivational emphasis on helping others and selflessness. Then, we found that male students had higher levels of Power (p < 0.001) and Achievement (p = 0.031), while female students outscored male students in Benevolence (p = 0.005) and Security (p = 0.006). Year of study showed no statistically significant difference. CONCLUSIONS Nursing students express high levels in hetero-directed values. Male nursing students, although they choose a stereotypically feminine profession, outscored females in stereotypical masculine values such as dominance and success. This is the first study that describes the personal value profile of undergraduate nursing students, according to the Theory of Basic Human Values, and it is a starting point for future research. CONCLUSIONS Nursing educators might want to consider the findings from this study while guiding students in developing awareness for their personal values.