Objectives Teaching involves multiple performance situations, potentially causing psychosocial stress. Since the theater-based improvisation method is associated with diminished social stress, we investigated whether improvisation lessened student teachers' stress responses using the Trier Social Stress Test (TSST; preparatory phase, public speech, and math task). Moreover, we studied the influence of interpersonal confidence (IC)the belief regarding one's capability related to effective social interactionson stress responses. Methods The intervention group (n = 19) received a 7-week (17.5 h) improvisation training, preceded and followed by the TSST. We evaluated experienced stress using a self-report scale, while physiological stress was assessed before (silent 30-s waiting period) and during the TSST tasks using cardiovascular measures (heart rate, heart rate variability [HRV]), electrodermal activation, facial electromyography (f-EMG), and EEG asymmetry. Hypothalamus-pituitary-adrenal (HPA-axis) reactivity was assessed through repeated salivary cortisol sampling. Results Compared to the control group (n = 16), the intervention group exhibited less f-EMG activity before a public speech and higher HRV before the math task. The low IC intervention subgroup reported significantly less stress during the math task. The controls showed a decreased heart rate before the math task, and controls with a low IC exhibited higher HRV during the speech. Self-reported stress and cortisol levels were positively correlated during the post-TSST preparatory phase. Conclusions These findings suggest that improvisation training might diminish stress levels, specifically before a performance. In addition, interpersonal confidence appears to reduce stress responses. The decreased stress responses in the control group suggest adaptation through repetition.