PURPOSE:To examine changes in macular ganglion cell layer (GCL) thickness over five years in a community-based birth cohort of children without ophthalmic disease. METHODS:This prospective study included 660 children without ophthalmic disease from the Copenhagen Child Cohort 2000 Eye Study. Optical coherence tomography (OCT) scans were analyzed for seven macular GCL thickness and volume measurements. Associations with age, sex, axial length, and OCT signal strength were evaluated. Additionally, GCL thickness was measured in eyes with optic disc drusen (ODD; n = 16) and compared with non-ODD eyes. RESULTS:From around age 11 to 16, macular GCL thickness in boys increased by approximately 1.6%, with statistical significance in six of seven predefined measurements. In girls, GCL thickness was stable or slightly decreased (-0.4%), with significance in four of seven measurements. Baseline axial length (AL) differed significantly between sexes (23.47/22.89 mm in boys/girls, p < 0.0001), as did axial growth between visits (ΔAL, 0.19/0.42 mm in boys/girls, p < 0.0001). No consistent associations were found between AL and GCL thickness or longitudinal changes thereof. After adjusting for potential confounders, four of seven GCL measurements increased significantly with age in boys, while no age-related changes were observed in girls. No significant differences were found in baseline GCL thickness or longitudinal change in GCL between eyes with or without ODD. CONCLUSIONS:GCL thickness remained relatively stable between ages 11 and 16, with small but statistically significant differences between sexes.