Background: University education is undergoing a paradigm shift towards active methodologies, such as virtual reality and training videos, which have proven to be valuable resources, especially in the health sciences. The scarcity of existing research on the topic prompted us to conduct this study, which seeks to measure the knowledge gained from the aforementioned tools by users, their level of satisfaction with them, and their perceived utility. Methods: This is a quasi-experimental intervention study analysing the impact of virtual objects as learning resources for undergraduate nursing students. Results: Fifty-four participants completed the training, yielding highly significant differences between their mean scores, with a high statistical power and a large effect size. A total of 85.46% of participants confirmed that the virtual resources helped them considerably to empathise with the experiences of trans people in healthcare settings. Students were comfortable using the virtual resources, very satisfied with the methodology employed, and would recommend the training received. Conclusions: University teaching must adapt to meet the current legislations and changing health needs of society, and teaching staff must be prepared to implement new active teaching methodologies that make learning a more dynamic process. Considering these results, our study serves as a guide for other nursing educators who seek to promote inclusive healthcare regarding gender diversity. This study is not registered.