Spondylolysis and spondylolisthesis are common findings in adolescent athletes (AA), potentially leading to load-dependent pain and segmental spine instability. In diagnostics, supine MRI is frequently used to visualize the lumbar spine structures. However, upright MRI procedures have been shown to be of clinical value in the detection of load-dependent changes in parameters. PURPOSE: To determine the feasibility of upright compared to supine MRI measurements to determine characteristics of the lumbar spine in AA with spondylolisthesis. METHODS: Ten AA (n=10; m/f: 4/6; 14.5±1.7y; 163±7cm; 52±8kg) from various sports, diagnosed with spondylolisthesis grade I-II Meyerding confirmed by x-ray in standing lateral view, were included. Open low-field MRI images (0.25 Tesla) in upright (82°) and supine (0°) position were evaluated by two observers. Medical imaging software was used to measure the anterior translation (AT, mm), lumbosacral joint angle (LSJA, °) and lordosis angle (LA, °). Reliability was analyzed by the intra-rater correlation coefficient (ICC) and standard error of measurements (SEM). RESULTS: Due to motion artifacts during upright position, measures of three participants had to be excluded. Between observers, AT ranged from 4.2±2.7mm to 5.5±1.9mm (ICC=0.94, SEM=0.6mm) in upright and from 4.9±2.4mm to 5.9±3.0mm (ICC=0.89, SEM=0.9mm) in supine position. LSJA varied from 5.1±2.2° to 7.3±1.5° (ICC=0.54, SEM=1.5°) in upright and from 9.8±2.5° to 10±2.4° (ICC=0.73, SEM=1.1°) in supine position. LA differed from 58.8±14.6° to 61.9±6° (ICC=0.94, SEM=1.19°) in upright and from 51.9±11.7° to 52.6±11.1° (ICC=0.98, SEM=1.59°) in supine position. CONCLUSIONS: Determination of AT and LA showed good to excellent reliability in both, upright and supine position. In contrast, reliability of LSJA had only moderate to good correlation between observers and should therefore be interpreted with caution. However, motion artifacts should be taken into consideration during upright imaging procedures.