This study aimed to examine the effects of aspirin on the growth and cardiomyocyte differentiation grade of bone marrow mesenchymal stem cells (BMMSCs). BMMSCs were divided into five differentiation groups with different concentrations of aspirin (0 mM, 0.5 mM, 1 mM, 2 mM, or 5 mM), and a undifferentiated control group. Cell growth was measured by cell proliferation, apoptosis assays and DNA cycle analysis. The differentiation grade of BMMSC-derived cardiomyocyte-like cells was examined by measuring the levels of cardiac-specific proteins with cyto-immunofluorescence staining, flow cytometry, and Western blotting. Electrophysiological analyses were performed by patch-clamp experiments and calcium transients were measured by a laser scanning confocal microscope. Cell proliferation decreased as the concentration of aspirin increased. Cell apoptosis increased with increasing aspirin concentration. DNA replication was inhibited in the high dose-aspirin group compared to the low dose-or non-aspirin groups. The number of a-myosin heavy chain (alpha-MHC) and cardiac troponin I (cTnI) positive cells, cardiac troponin T (cTnT) and connexin 43 (Cx43) positive rates, expression levels of Cx43, Nkx2.5, GATA4 and beta 1 adrenoceptor increased with increasing aspirin concentration. No sarcomeric cross-striations, spontaneous or induced beating activity or action potentials was observed in each group. Calcium transients were measured in small number cells in 2mM aspirin group, but the features are atypical. Consequently, aspirin inhibits proliferation and survival of BMMSCs and enhances cardiomyocyte differentiation of BMMSCs.