To compile and statistically summarize quantitative evidence on the acute effects of resistance training sessions on muscle glycogen concentration, a systematic search was conducted on Pubmed, Web of Science, and Scopus databases up to 28th July 2024. Twenty studies including 168 male and 12 female participants were eligible. A multilevel, random-effects meta-analysis was used to calculate the overall mean difference (MD) with a 95% confidence interval (CI) and prediction interval (PI). The model (28 effect sizes across 20 clusters) revealed a significant glycogen decrease (MD = −104.3; 95% CI: −137.6 to −71.0; PI: −244.4 to 35.7; p < 0.001). Meta-regression showed greater depletion with more sets (Estimate = −11.2; 95% CI: −18.0 to −4.3; p = 0.001) and longer session duration (Estimate = −1.3; 95% CI: −2.3 to −0.3; p = 0.009), but less with higher intensity (Estimate = 2.88; 95% CI: 1.2 to 4.5; p = 0.0006). Subgroup analysis showed greater depletion with varied intensity (MD = −162.9) versus fixed (MD = −82.5), and in untrained (MD = −113.0) versus trained participants (MD = −101.3). A single resistance training session depletes glycogen in the vastus lateralis muscle, with depletion influenced by training intensity, session duration, number of sets within session and training status.