The current study examines the association between the mother-caregiver relationship and the psychological well-being of 533 mothers who transitioned their children to a non-familial licensed childcare center. Maternal well-being, mother-caregiver relationship quality, and maternal parenting stress were assessed. Hierarchical linear regressions were conducted to examine the associations between independent variables (i.e., maternal demographic data, parenting stress, and maternal reports of supportive caregiver relationship) and maternal well-being as a dependent variable. The results show that together, the independent variables account for 31% of variation in maternal well-being. Findings suggest that a supportive caregiver relationship is a significant factor in contributing to maternal well-being. There is a call to recognize mothers transitioning their children to non-familial care as a special population for consideration by teachers, caregivers, and infant/child mental health professionals working in childcare settings. The current research bolsters discussion on mother-caregiver relationships to focus on how professionals in childcare settings can best support mothers in this unique transition to non-familial care for their children.