Hills, SP, Barrett, S, Busby, M, Kilduff, LP, Barwood, MJ, Radcliffe, JN, Cooke, CB, and Russell, M. Profiling the post-match top-up conditioning practices of professional soccer substitutes: An analysis of contextual influences. J Strength Cond Res 34(10): 2805–2814, 2020—Soccer practitioners implement “top-up” conditioning sessions to compensate for substitutes' limited match-play exposure. Although perceived to be valuable for reducing injury risk and augmenting positive physical adaptations, little research has considered the demands of post-match top-up training. To quantify post-match top-up responses, 31 professional soccer players wore 10 Hz microelectromechanical systems after 37 matches whereby they were selected in the match-day squad as substitutes (184 observations; 6 ± 5 observations·player−1). Linear mixed models and effect sizes (ES) assessed the influence of contextual factors on 23 physical performance variables. Top-ups lasted 17.13 ± 7.44 minutes, eliciting total and high-speed distances of 1.7 ± 6.2 km and 0.4 ± 1.7 km, respectively. Each contextual factor (i.e., position, substitution timing, match location, result, time of day, stage of the season, and fixture density) influenced at least 4 of the dependent variables profiled (p ≤ 0.05). Top-up duration; total, moderate-speed, and low-speed distance; and the number of repeated high-intensity efforts were greater for unused vs. used substitutes (ES: 0.38–0.73, small to moderate). Relative to away matches, home top-ups elicited heightened total, low-speed, and high-speed distances, alongside more moderate-speed accelerations and decelerations, and repeated high-intensity efforts (ES: 0.25–0.89, small to moderate). Although absolute and relative running distances were generally the highest when the fixture density was low, the greatest acceleration and deceleration demands were observed during the most congested fixture periods. Late-season top-ups typically elicited lower absolute physical responses than early and mid-season sessions. These data provide important information for practitioners when considering the aims and design of substitute top-up conditioning sessions, particularly with reference to contextual influences.