Human milk is the preferred choice for infant feeding. When a sick or premature infant’s own mother’s milk is unavailable, donor human milk is becoming more widely used. Many potential milk donors do not live within close proximity to the 10 North American not-for-profit milk banks. Transporting milk via commercial carriers can be inconvenient and costly for recipient banks. A network of donor human milk depots is one practical way to increase the quantity of available donor human milk. This article provides guidelines and practical suggestions for establishing a donor human milk depot.