Although co-teaching is increasingly recognized as a valuable approach in inclusive education, comprehensive studies that examine the impact of targeted professional development on enhancing teachers’ capabilities in this area are lacking. The limited research on this topic fails to provide clear evidence of the effectiveness of such training programs and their potential benefits for special education and general education teachers. Addressing this issue, this study investigates the effectiveness of a proposed professional development training program designed to elevate co-teaching and collaboration skills among in-service teachers. A one-group quasi-experimental design was used, and a convenience sampling of 32 female teachers, comprising 16 special education and 16 general education teachers, participated in the program consisting of 14 intensive sessions. Pre- and post-intervention assessments, comprising a co-teaching test and a collaboration skills scale, were administered. Results demonstrate significant benefits of the training program, with participants showcasing improved performance in the co-teaching post-test and collaboration scale. Special education teachers notably outperformed their general education counterparts in co-teaching skills. However, the analysis did not reveal any statistically significant differences between teachers of students with learning disabilities and teachers of general education regarding their performance on the collaboration skills scale. The study concludes with recommendations and future research directions, offering insights to further advance co-teaching practices and collaboration skills among educators.