The purpose of this study was to examine the levels of self-esteem among pre-service teachers in Turkey and Iran who were studying in the Physical Education and Sports Teaching Departments at their respective countries. The study's participant group consisted of 80 Turkish and 111 Iranian students. The Coopersmith Self-Esteem Inventory was used for this cross-cultural comparative research approach. A two-way analysis of variance was used to examine the data. When the self-esteem scores of pre-service teachers were examined, it was shown that pre-service teachers in both Turkey and Iran scored higher than the national average. Although the mean self-esteem ratings of pre-service teachers in Turkey were found to be higher than those of pre-service teachers in Iran, it could be claimed that the self-esteem levels of pre-service teachers did not differ according to the culture they lived in, in other words, they were at a similar level. Similarly, it was discovered that self-esteem levels did not differ significantly by gender, grade level, perceived level of academic accomplishment, or cultural characteristics. One of the key elements of human personality is the sense of self-esteem which influences both the level of self-acceptance and self-awareness. Therefore, it is of paramount importance to develop these skills and the psychophysical characteristics in the process of upbringing and education.