Objectives Low-intensity ultrasound (LIUS) has been shown to enhance bone and cartilage regeneration from stem cells. The ease of its incorporation makes it an attractive mechanical stimulus for not only osteogenesis and chondrogenesis, but also cardiomyogenesis. However, to date, no study has investigated its effects on cardiomyogenesis from embryonic stem cells. Methods In this study, murine embryonic stem cells were differentiated via embryoid body formation and plating, and after 3 days they were subjected to daily 10 minutes of LIUS treatment with various conditions: (1) low-pulsed (21 mW/cm2, 20% duty cycle), (2) low-continuous, (3) high-pulsed (147 mW/cm2, 20% duty cycle), and (4) high-continuous LIUS. Results Low-pulsed and high-continuous LIUS had improved beating rates of contractile areas as well as increased late cardiac gene expressions, such as α- and β-myosin heavy chain and cardiac troponin T, showing its benefits on cardiomyocyte differentiation. Meanwhile, an early endodermal marker, α-fetoprotein, was significantly attenuated after LIUS treatments. Conclusions With these observations, it is demonstrated that LIUS simulation could enhance cardiomyogenesis from embryonic stem cells and increase its selectivity toward cardiomyocytes by reducing spontaneous differentiation.