Cells cultured play an important role in organism-resource protection and functional identification of genes, as well as mechanism exploration and prevention of animal disease. However, tissue cells that can survive for a long time are limited at present in marine shellfish. In this study, primary cultures of heart cells from the scallop were started using an explant method, and a primary culture system-which can keep the cells alive for a long time -was established using an optimizing-medium method. Effects of three basic media (L-15, M199, and L-15+M199) on the number of migrated cells from the explants and cell survival time were compared; the L-15 medium was verified to be the optimal basic medium for heart cells. Furthermore, the optimal supplemental-factor combination for heart cells was the L-15 medium supplemented with 5% FBS, 50 mmol/L taurine and 6 mmol/L Ca²⁺ (orthogonal experiment of three factors and three levels), in which cardiomyocytes could survive for 2 months . In this study, the results show that low concentration serum (5% FBS) was most favorable for the migration of cells from the tissue mass. With an increase in FBS concentration, the cell-migration ability was weaker, and the survival time of primary cells became shorter. After the addition of taurine, the survival time of primary cells was significantly prolonged. However, the Ca²⁺ (2 mmol/L, 4 mmol/L, and 6 mmol/L) had no significant effect on heart-cell cultures. Perhaps the additive taurine regulated intracellular Ca²⁺ to a suitable level, resulting in the addition of Ca²⁺ without significant improvement in the primary culture. In this primary culture, most cells were cardiomyocytes; part of the cardiomyocytes beat at regular intervals within a short time, and cardiomyocytes and myotubes were formed in part of the region. This phenomenon was also reported in the primary culture of heart cells of This study provides a useful foundation for further studies on basic biology and functional genes.