Abstract: Objective To seek a simple and effective approach through probing method about primary culture and purification of neonatal rat cardiac myocytes. Methods Twenty neonatal rats were randomly divided into 2 groups. Group A: the cardiac myocytes were gained by means of enzymic digestion and tubularis-blowing; Group B: the cardiac myocytes were gained by means of enzymic digestion and rotator-stirring. The cardiac myocytes in both groups were purified by means of differential attachment technique and BrdU inhibition and were identified with immunofluorescence staining. Viability was assessed with trypan blue staining and pulsation was assessed under microscope. Results The rates of cardiac myocyte’s viability, pulsation and purity in group A were (89.90±2.92)%, (91.30±2.00)% and (94.90±1.79)% respectively; The corresponding rates in group B were (94.70±2.31)%, (95.00±1.24)% and (95.60±1.43)% respectively. The rates of cardiac myocyte’s viability, pulsation in group B were significantly higher than that of group A, which was statistically different (\P>0.05). The rate of cardiac myocytes purity in group B was a bit higher than group A, but which wasn’t statistically different (\P<0.05). Fibroblasts cultured with BrdU grew weaker and less than those without BrdU. Conclusion The method of rotator-stirring and enzymic digestion was better than that of tubularis-blowing and enzymic digestion, which helped to gain cardiac myocyte effectively. BrdU helped to purify cardiac myocytes.