Background Cardiomyocyte transplantation for the therapy of myocardial ischaemia is being paid close attention. However, how the microenvironment controls the differentiation of transplanted bone marrow stromal cells (BMSCs) is unknown. Endothelin-1 (ET-1), a cytokine, increases during myocardial infarction, but it is not known whether ET-1 is responsible for the fate of transplanted BMSCs. In the present study, we investigated the effects of ET-1 on differentiation and maturation of induced rabbit BMSCs, in vitro, to elucidate the cellular biological mechanisms. Methods The proliferation of BMSCs isolated from femur of rabbits was induced by ET-1 only, by 5-azacytidine (5-aza) or ET-1 combined with 5-aza. After 4 weeks of induced culturing, the differentiation rate and the diameter of induced myocyte like cells were estimated and the expressions of GATA-4 protein and phosphorylation level were assayed by Western-blot, RT-PCR analysis of β-myosin heavy chain (MHC). mRNA expression, levels of troponin-I by immunohistochemical staining and ultrastructure of induce-cultured BMSCs were also determined. Results By induction with ET-1 and 5-aza, mean cell diameter of induced BMSCs was larger than induced with 5-aza [(6.26±0.22) μm cf (5.29±0.19) μm] (P < 0.001). There was no difference in rate of differentiation of myocyte like cells between the groups induced with 5-aza and ET-1 combined with 5-aza [(29.82±0.23) % cf (29.94±0.18) %] (P > 0.05). The expressions of GATA-4 protein and phosphorylation were enhanced significantly in groups induced with ET-1 combined with 5-aza (P < 0.05). In the group induced with ET-1 combined with 5-aza, expression of β-MHC mRNA was higher than control [(0.122±0.008) cf (0.022±0.003)] (P <0.01), and more troponin-I positive cells were also detected in this group. Differentiated BMSCs showed formations of myofilaments and primitive sarcomere, i.e., morphological characteristics of myocyte like cells. Conclusions This study suggests that induced culturing of BMSCs by ET-1 combined with 5-aza can express cardiomyocytic characteristics whereas ET-1 alone could not induce BMSCs to differentiate to myocyte like cells. ET-1 upregulates the expression of GATA-4 protein and phosphorylation level of induced BMSCs, and rapidly promotes the differentiation and maturation of myocyte like cells from BMSCs. However, how the microenvironment controls the differentiation of transplanted bone marrow stromal cells (BMSCs) is unknown. Endothelin-1 (ET-1), a cytokine, increased during myocardial infarction, but it is not known whether ET-1 is responsible for the fate of transplanted BMSCs. In the present study, we investigated the effects of ET-1 on differentiation and maturation of induced rabbit BMSCs, in vitro, to elucidate the cellular biological Methods The proliferation of BMSCs isolated from femur of rabbits was induced by ET-1 only, by 5-azacytidine (5-aza) or ET-1 combined with 5-aza. After 4 weeks of induced culturing, the differentiation rate and the diameter of induced myocyte like cells were estimated and the expressions of GATA-4 protein and phosphorylation level were assayed by Western-blot, RT-PCR analysis of β-myosin heavy chain (MHC) vels of troponin-I by immunohistochemical staining and ultrastructure of induce-cultured BMSCs were also determined. By By induction with ET-1 and 5-aza, mean cell diameter of induced BMSCs was larger than induced with 5-aza [(6.26 ± 0.22 ) were no difference in rate of differentiation of myocyte like cells between the groups induced with 5-aza and ET-1 combined with 5-aza [(29.82 ± 0.23 )% cf (29.94 ± 0.18)%] (P> 0.05). The expressions of GATA-4 protein and phosphorylation were significantly increased in groups induced with ET-1 combined with 5-aza (P <0.01), and more troponin-I positive cells were also detected in ((0.122 ± 0.008) cf (0.022 ± 0.003)] with ET-1 combined with 5-aza this group. Differentiated BMSCs showed formations of myofilaments and primitive sarcomere, ie, morphological characteristics of myocyte like cells. Conclusions This research suggests that induET-1 upregulates the expression of GATA-4 protein and phosphorylation level of induced BMSCs , and rapidly promotes the differentiation and maturation of myocyte like cells from BMSCs.