Differentiation of brown adipose-derived stem cells into cardiomyocyte-like cells is regulated by a combination of low 5-azacytidine concentration and bone morphogenetic protein 4.

Adipose-derived stem cells (ADSCs) could be an ideal candidate for seed cells to regenerate damaged heart tissue. This study examined and compared the cardio-myogenic differentiation efficacy of neonatal rat brown ADSCs (rbADSCs) treated with either 5-azacytidine (5-AZA), bone morphogenetic protein 4 (BMP4), or lower doses of both molecules. Briefly, by investigating the protein expression of cardiac-specific markers (i.e., cardiac troponin-I, α-sarcomeric actinin, sarcoplasmic reticulum Ca2+-ATPase, and connexin 43), our data indicated that rbADSCs could be differentiated into cardiomyocyte-like cells by all three treatments. By quantitatively measuring the number of cells with positive staining for the above markers, we found that the low-dose combined treatment showed higher differentiation efficiency compared to standard dose 5-AZA and BMP4 treatment. Similarly, the expression levels of these proteins as determined by western blotting were higher in the low-dose combination group than in the standard dose 5-AZA and BMP4 groups. Also, the combined strategy maintained the decreased cell viability caused by cytotoxicity of 5-AZA, probably through reducing the ratio of apoptotic rbADSCs. Furthermore, the extracellular regulated protein kinase (ERK) signaling pathways participate in the differentiation process, but the observed effects between the BMP4 and 5-Aza treatments are quite different. IJCEP