Cell transplantation which aims to introduce healthy myogenic cells into the myocardium of the diseased heart is an important tool in myocardial regeneration therapy. The differentiation potential of bone marrow mesenchymal stem cells (BM-MSCs) can be key role for this purpose. The present study aimed to evaluate cardiomyocyte gene expression profile of 5-azacytidine treated rat BM-MSCs. 5-Aza treated BM-MSCs started to form cell colonies and especially these colonies stained with desmin and cardiac troponin-T. qRT-PCR results indicate that the expression level of cardiomyocyte specific genes especially α- cardiac actin, Mef-2b, GATA-4 were increased in 5 μM 5-Aza treated BM-MSCs. According ELISA results, nanogram levels of cardiac troponin-I was measured especially 5 μM 5-Aza treated cells.