Nesprin-1 has key roles in the process of mesenchymal stem cell differentiation into cardiomyocyte-like cells in vivo and in vitro.

Abstract

The aim of the present study was to investigate the expression of nesprin-1 protein in MSCs and its effects on the differentiation of rat bone-marrow mesenchymal stem cells (MSCs). Surface-associated antigens of MSCs were detected by flow cytometry. MSC differentiation was induced by treatment with 10 μmol/l 5-azacytidine. Sprague-Dawley rats were anesthetized prior to thoracotomy and subsequent ligation of the left anterior descending coronary artery to establish a model of myocardial infarction. Two weeks following myocardial infarction, DAPI-marked MSCs were injected into the infarcted region in the experimental group, while DMEM was injected into the infarcted region of the control group. Characteristics of the putative cardiac-myogenic cells were evaluated using immunohistochemical and immunofluorescent analysis. The messenger RNA expression levels of cardiac-myogenic specific genes; desmin, α-actinin and cardiac troponin I (cTnI) were detected by reverse transcription quantitative polymerase chain reaction. The expression of nesprin-1 protein in MSCs was identified by immunofluorescence and western blot analysis, prior to and following MSC differentiation. Following differentiation, the MSCs appeared spindle-shaped with irregular processes and were positive for CD90 and CD29, but negative for CD45. Cardiomyocyte-like cells were positive for desmin, α-sarcomeric actin and cTnI. Nesprin protein was detected in the nuclear membrane via immunofluorescence, and following MSC differentiation into cardiomyocyte-like cells, the expression of nesprin protein was significantly higher (*P=0.03<0.05). The results of the present study indicated that MSCs may be differentiated in vitro and in vivo into cells with characteristics commonly attributed to cardiomyocytes. Cardiomyocyte-like cells cultured from bone marrow sources may be potentially useful for repairing the injured myocardium. The results also suggested that, consistent with the results of previous studies, the expression of nesprin-1 protein was higher during the differentiation process of MSCs and may have an important role in mediating MSC differentiation. Elucidation of the role of nesprin-1 in MSC differentiation may aid in the development of novel therapies for the treatment of myocardial ischemia and nesprin-1 genetic deficiencies.