Umbilical cord blood collection increases the life expectancy of end-stage or chronically ill patients. The knowledge and attitude of health professionals are essential to the success of this attempt. This inquiry aimed to evaluate the effect of applying a structured teaching program on knowledge and attitude concerning umbilical cord blood collection and its barriers among maternity nurses. A quasi-experimental research design was used. It was carried out in Obstetrics and Gynaecological departments (antenatal, postnatal, and operating room) at Banha University Hospital. Based on a convenient sampling method, 89 maternity nurses at Banha University Hospital were chosen. To gather the data, two instruments were utilized: a) An organized interview questionnaire sheet, including two sections of socio-demographic features of maternity nurses and knowledge of maternity nurses concerning umbilical cord blood collection and its barriers; b) a modified Likert scale to assess the nurses' attitudes regarding umbilical cord blood collection and its barriers. The results showed that a minority of the nurses (11.2%) had good knowledge about umbilical cord blood collection and its barriers pre-intervention, which increased to (86.5%) post-intervention; however, only one-fifth of nurses (20.0 %) had a positive attitude of umbilical cord blood collection and its barriers pre-intervention which increased to (89.9%) post-intervention. Additionally, a highly direct correlation between total knowledge and total attitude score pre-and post-intervention was noticed. Structured teaching program regarding umbilical cord blood collection significantly improved nurses' knowledge and attitude regarding umbilical cord blood collection and its barriers. As a recommendation, appropriately designed in-service training programs concerning umbilical cord blood banking and its barriers must be founded to improve nurses' knowledge and attitude to suit newly developed concepts in care.