5-Azacytidine Promotes Differentiation of Bone Marrow Derived Mesenchymal Stem Cells into Troponin Positive Cardiomyocyte‑Like Cells in Vitro.

Objective: The aim of this work is to differentiate Mesenchymal stem cells derived (MSCs) from bone marrow obtained from human adult into cardiomyocyte using 5-azacytidine in vitro.Background: MSCs are considered a promising tool for cellular therapy because of their capability for selfrenewing and their multipotency.Owing to the limited ability of the injured heart to regenerate cardiomyocytes.Cardiomyocyte transplantation has been advocated as a possible treatment for heart failure.Their other benefits on a myocardiac infarct is owed to neo-vascularization, because they can differentiate also into endothelial cells.Methods: MSCs were cultured from bone marrow aspirate and detected morphologically and by flowcytometric analysis of surface markers CD44 and CD 34 then differentiated to cardiomyocytes by using 5azacytidine .The cardiomyocytes were detected morphologically and by flowcytometric analysis of Vimentin and Troponin and immunofluorescence analysis for Cardiac alpha tropomyosin Results: MSCs can be isolated from bone marrow and assumed the typical fibroblastoid morphology and reach 80% -90% confluence at about 9 days.They express CD44 and are negative for CD 34 .MSCs induced with 5azacytidine show morphological changes consistent with cardio myocytes as compared to the symmetric morphologies of the uninduced cells, as shown by inverted microscope.Induced cells showed positive expression of troponin and vimentin and positive cardiac alpha tropomyosin.. Conclusion:MSCs can be successfully isolated from bone marrow aspirate and can be differentiated into cardiomyocytes.