The authors studied quality control procedures at human milk banks and nutritional profiles of 909 milk samples (from 195 donors, aged 15 to 45 years) from banked human milk (BHM) in Brasília, Brazil. Number of donations per donor ranged from 1 to >10 that consisted mostly of mature milk (90.9%) with a mean total energy of 529±85 kcal/L and a mean total lipid of 22.7 g/L ±13.2. Microbiological quality (titrable acidity—Dornic, °D) was suitable for infant feeding in 99.2% of samples (≤ 8°D), ranging from 2°D to 8°D (mean 4.8 ± 1.4°D). Most BHM (98.1%) samples were dispensed to inpatient infants (1–7 days) diagnosed with respiratory distress (30.1%), prematurity (20.7%), metabolic distress (16.0%), jaundice (14.4%), bacterial infection (6.0%), pneumonia (3.3%), congenital cardiac distress (2.2%), or other conditions (6.2%). Well-motivated mothers and trained staff are serving and sustaining an important life-saving network with long-lasting impact on public health. J Hum Lact. 22(3):335–339.