Culture and identification of cardiomyocytes in neonatal rats

AIM:To develop a method of primary culture of neonatal rat cardiomyocytes with high survival cell rate and high purity. METHODS: Hearts of 1 to 2dayold neonatal Sprague Dawley rats were removed under sterile conditions with 0.25% trypsin and 02% collagenase type II at 37℃ under the conditions of graded digestion myocardial tissue. Differential adherence purification was applied. The time was extended to 90 min and the medium was changed after 48 h. RESULTS: Adherent growth of myocardial cells was observed at 24 h and pseudopodia growth at 2-3 days. Myocardial cells formed cell clusters and the synchronous beat frequency was about 80-130 times/min with survival rate of (93.9±2.8)% and purity of (91.9±2.2)%. CONCLUSION: We developed a method to culture myocardial cells with high survival cell rate and high purity.