Measurements of muscle-tendon unit passive mechanical properties are often used to illustrate acute and chronic responses to a training stimulus. The purpose of this study was to quantify the inter-session repeatability of triceps surae passive stiffness measurements in athletic and non-athletic populations, with the view to discussing its usefulness both as a muscle-tendon profiling tool and a control measure for studies with multiple data collection sessions. The study also aimed to observe the effects of quiet standing on passive stiffness parameters. Twenty-nine men (10 cyclists, nine triathletes, 10 controls) visited the laboratory on three separate occasions, where passive stiffness tests were carried out using an isokinetic dynamometer and B-mode ultrasound. Participants were fully rested on two of the sessions and subjected to 20min of quiet standing in the other. The passive stiffness assessment generally showed only moderate inter-session repeatability but was still able to detect inter-group differences, with triathletes showing higher passive stiffness than cyclists (p<0.05). Furthermore, quiet standing impacted passive stiffness by causing a reduction in ankle joint range of motion, although mechanical resistance to stretch in the muscle-tendon unit at a given joint angle was relatively unaffected. These findings show that passive stiffness assessment is appropriate for detecting inter-group differences in the triceps surae and even the effects of a low-intensity task such as quiet standing, despite showing some inter-session variation. However, the inter-session variation suggests that passive stiffness testing might not be suitable as a control measure when testing participants on multiple sessions.