Background: Mothers who applied kangaroo care for their premature neonates have more milk expression than those who didn't. The study aimed to determine the effect of kangaroo care applied by mothers of premature neonates on the amount of maternal milk expression. Settings: The study was conducted at three Neonatal Intensive Care Units; maternity University Hospital at shatby, Alexandria university children's hospital at Smouha and in El-Ramel hospital for children (wingat). Subjects: A convenient sample of 60 mothers who have admitted premature neonates in the previously mentioned settings comprised the study subjects. Mothers who are visiting their premature neonates regularly (five days a week with a duration of three weeks). Tools: Two tools named Socio-Demographic characteristics of mothers and clinical data of the premature neonates structured interview schedule as well as Maternal milk expression record were used for data collection. Results: significant difference was shown between the three studied weeks for the mothers in the study group (Fr=12.0, p=0.002). Where more than two third of mothers (70%) expressed 500 ml and more at the end of third week compared to those mothers who expressed breast milk at the end of second week (63.3%) and first week (33.3%). Conclusion: It was concluded from this study that practice of kangaroo care affecting the amount of expressed breast milk where mothers who applied it expressed more amount of breast milk than those who didn't. Recommendation: It was recommended that kangaroo care should be considered an essential method for caregivers regarding their neonates as early as possible and should be included in strategies and policies of hospital.