Cortisol is the primary output of the hypothalamic-pituitary-adrenal (HPA) axis and is central to the human biological stress response, with wide-ranging effects on physiological function and psychiatric health. In both humans and animals, cortisol is frequently studied as a biomarker for exposure to environmental stress. Relatively little attention has been paid to the possible role of genetic variation in heterogeneity in chronic cortisol, in spite of well-studied biological pathways of glucocorticoid function. Using recently developed technology, hair samples can now be used to measure accumulation of cortisol over several months. In contrast to more conventional salivary measures, hair cortisol is not influenced by diurnal variation or transient hormonal reactivity. In an ethnically and socioeconomically diverse sample of 1 070 child and adolescent twins and multiples from 556 unique families, we estimated genetic and environmental influences on hair concentrations of cortisol and its inactive metabolite, cortisone. We identified sizable genetic influences on cortisol that decrease with age, concomitant with genetic influences on cortisone that increase with age. Shared environmental influences on cortisol and cortisone were modest and, for cortisol, decreased with age. Twin-specific, non-shared environmental contributions to cortisol and cortisone became increasingly correlated with age. We find some evidence for sex differences in the biometric contributions to cortisol, but no strong evidence for main or moderating effects of family socioeconomic status on cortisol or cortisone. This study constitutes the first genetic study of hormone concentrations in human hair, and provides the most definitive characterization to-date of age and socioeconomic influences on hair cortisol.