Background: Survival rate of premature neonates has increased due to the technological advances in medical care. Developmental care interventions may help preterm neonates cope better with the environment of the Neonatal Intensive Care Unit and provide healthy developmental outcomes.

The study was aimed: to evaluate the effect of developmental supportive care training program on nurses’ practice regarding behavioral response of premature neonates.

Subjects and method: A convenience sampling of fifty nurses who were responsible for providing nursing care for premature neonates was included in the study, also, all premature neonates who receiving care were included in the study, Three tools were used to collect data: Structured interview schedule, Nurses practice of Developmental Supportive Care for premature neonates using an observational checklist, Observational checklist to assess nurse's practice regarding behavioral response of premature neonates.

The results: revealed that there was statistically significant difference of levels of total nurses 'knowledge between before, immediate, and after one month of a supportive developmental care training program.

The study was concluded: that developmental supportive care had a positive effect on nurses’ knowledge and practice as well as their perceived self-competence regarding behavioral response of premature neonates as evidenced improved their oxygen saturation, regulate heart rate and respiration.

The study was recommended: that application of administrative policy and procedures for nursing practice of developmentally supportive care for high-risk neonates are needed and incorporate neonatal intensive care units design.