Student training requires specific laboratories for vaccination practice, which are usually limited, and even professionals' continuing education regularly lacks proper care. Thus, new methodologies, concepts, and technologies, such as software-based simulations, are in highly demand. This work aims to develop a 3D virtual environment to support teaching activities in the vaccination room. The software-based simulation must contribute positively to teaching considering a variable set of scenarios. We applied the design science research method to guide the work. First, the concepts and opportunities were raised, which we used to build the simulation (ie, the proposed technological artifact). The development was assisted by a specialist, in which we sought to create a vaccination room according to Brazilian standards. The artifact evaluation was achieved in 2 stages: (1) an evaluation to validate the design with experts through the Delphi method; and (2) a field evaluation with nursing students to validate aspects of usability (System Usability Scale [SUS]) and technology acceptance and use (Unified Theory of Acceptance and Use of Technology version 2). We built the simulation software using the Unity game engine. An additional module was also developed to create simulation scenarios and view the students' performance reports. The design evaluation showed that the proposed solution is adequate. Students' evaluations confirm good usability (SUS score of 81.4), besides highlighting Performance Expectation as the most positively influential factor of Behavioral Intention. Effort Expectancy is positively affected by younger users. Both evaluation audiences cited the high relevance of the proposed artifact for teaching. Points for improvement are also reported. The research accomplished its goal of creating a software-based simulation to support teaching scenarios in the vaccination room. The evaluations still reveal desirable improvements and user behavior toward this kind of technological artifact.