Many studies have noted that the bones of the human upper limb display bilateral asymmetry, commonly linking this asymmetry in external and internal morphology to handedness and lateralization. Few studies, however, have attempted to track asymmetry throughout ontogeny. This study assesses the ontogenetic development of cortical and trabecular bone asymmetry in the humerus. We predict that directional asymmetry in structural properties will emerge in concert with hand preference and increased activity levels during the juvenile period. Paired humeri from 57 individuals from the Norris Farms #36 archaeological skeletal collection ranging in age from neonate to adult were used in the current study. Cortical bone cross-sectional properties and three-dimensional trabecular bone structure were quantified from microcomputed tomography data. The results indicate significant absolute asymmetry in all measured cortical and trabecular bone variables across all ages. Trabecular bone displays significantly higher absolute asymmetry than cortical bone. Contrary to expectations, however, this study found very little evidence for significant directional asymmetry in humeral length and cortical or trabecular bone variables, except in adults. The presence of significant absolute asymmetry in all age groups, and the lack of significant directional asymmetry in almost all variables at all ages, suggests that structural differences due to higher levels of habitual loading in the dominant arm are overlain on a template of potentially significant existing asymmetry. Anat Rec, 301:1012-1025, 2018. © 2017 Wiley Periodicals, Inc.