Reliably eliciting acute stress repeatedly over time is of indispensable value for research into stress vulnerability and for developing interventions aimed at increasing stress resiliency. Here, we evaluated whether the Maastricht Acute Stress Test (MAST), a potent stress protocol that combines physical and psychosocial stress components, can be used to reliably elicit subjective and neuroendocrine stress responses multiple times. Sixty healthy undergraduate participants were exposed to the MAST on three occasions, with intervals of three-weeks and one-month in between sessions. Results showed no significant signs of habituation or sensitization to the MAST in terms of subjective or physiological (salivary alpha-amylase and cortisol) stress reactivity. Fifty-nine percent of the sample displayed a significant physiological stress response (i.e., cortisol) to two MAST exposures and 57% to every MAST exposure. This study demonstrates that the MAST can be used to repeatedly induce significant stress responses.