Background: The aim of this paper is to analyze the results of virtual reality (VR) antromastoidectomy simulation training and the transferability of the obtained skills to real temporal bone surgery. Methods: The study was conducted prospectively on a group of 10 physicians, and was composed of five VR simulation training sessions followed by live temporal bone surgery. The quality of performance was evaluated with a Task-Based Checklist (TBC) prepared by John Hopkins Hospital. Additionally, during every VR session, the number and type of mistakes (complications) were noted. Results: The quality of performance measured by the TBC increased significantly during consecutive VR sessions. The mean scores for the first and fifth sessions were 1.84 and 4.27, respectively (p < 0.001). Furthermore, the number of mistakes in consecutive VR sessions was gradually reduced from 11 to 0. During supervised surgery, all the participants were able to perform at least part of an antromastoidectomy, and the mean TBC score was 3.57. There was a significant strong positive correlation between the individual results of the fifth VR session and the individual results of supervised surgery in the operating room (rp = 0.89, p = 0.001). Conclusions: Virtual reality for temporal bone training makes it possible to acquire surgical skills in a safe environment before performing supervised surgery. Furthermore, the individual final score of virtual antromastoidectomy training allows a prediction of the quality of performance in real surgery.