Introduction: Prematurity is the most common cause of neonatal mortality and morbidity. Most preterm infants receive multiple packed red cell (PRC) transfusions during hospitalization. Meanwhile, multiple PRC transfusions can increase iron levels. However, to date, there is no consensus regarding iron supplementation in preterm who have received multiple PRC transfusions. The objective of this study is to determine the iron status of premature infants aged 28-32 weeks who have received multiple PRC transfusions and to make recommendations regarding iron supplementation. Methods: This study is a prospective cohort study of 70 preterm infants born at the Cipto Mangunkusumo Hospital in March 2021-May 2021. Iron profiles were examined at 1, 2, and 3 months of chronological age. Results: At 1 month chronological age, infants who received PRC transfusions >2 times had significantly higher iron profiles compared to subjects who received PRC transfusions 1-2 times only, and the total iron binding capacity (TIBC) levels were significantly lower in subjects who received PRC transfusions >2 times than infants who received PRC transfusions 1-2 times. The cut-off point for the total volume of PRC transfusion that causes iron overload status is ≥50 mL/kg. The median serum ferritin (Fe) levels in infants who received more than two PRC transfusions were 498.11 µg/L (range: 358-885.62 µg/L) at 1 month of age, 232.66 µg/L (range: 60.85-538.44 µg/L) at 2 months, and 42 µg/L (range: 40.1-168.63 µg/L) at 3 months. Conclusions: The results of this study can be used in limited health facilities that do not have the availability of iron profile examinations. Preterm infants who received more than two PRC transfusions had a higher iron profile and lower TIBC levels at 1 month of chronological age compared to those who received two or fewer transfusions. Preterm infants with PRC transfusion >2 times or ≥50 mL/kg exhibited iron overload by 1 month of chronological age; therefore, iron supplementation may be considered to be initiated at 2 months of chronological age. © 2025 National Neonatology Forum