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Shellfish with high economic value, aquaculture has become a pillar industry of China's marine aquaculture industry. In recent years, the mariculture shellfish massive death event occurs frequently, the urgent need to establish a stable shellfish cell culture system, from the study of pathogens and their pathogenic mechanisms of the cellular and molecular levels, so as to effectively detect, prevent the occurrence of diseases. The cells cultured easy breeding techniques of cell engineering to breed new varieties with strong resilience, but also to solve one of the means of disease control. Shellfish rich germplasm resources, the establishment of cell lines will contribute to the long-term preservation of genetic resources. In addition, with the increase in atmospheric pollution, shellfish as an indication of the sensitive biological role has become increasingly prominent. In this study, our northern coastal aquaculture shellfish C. farreri as the research object, cell culture studies, aims to establish a set of C. farreri tissue cells in vitro long-term culture method for immunological studies, the study of pathogen infection mechanism species conservation, genetics and breeding, environmental monitoring, a platform, Chlamys farreri cell line and eventually build the foundation. The Chlamys farreri myocardial tissue in primary cultured single-factor optimization results show that the basic medium L-15 to maximize promote explant cells to move out and a long time to maintain cells in vitro survival, L-15, M199, M199 less effective, followed by ; 5? S to maximize the promotion of the explant cells move out and a long time to maintain cells in vitro survival, 10? S, followed by 15% and 20? and S less effective. In the under basic medium L-15 ,5-10? S culture conditions, the cell average survival of about two weeks, some of the cells can be maintained 50d. Cardiomyocytes cultured in vitro with active contraction phenomenon, cell monolayers at the form myotubes and myofilament structure. Ultrastructural myofibrillar culture the 10d after myocardial cells are localized cultivate longer myocardial cells the sarcoplasmic reticulum swollen to several vacuoles. With culture, cell succinate dehydrogenase activity gradually weakened, and alkaline phosphatase activity is relatively stable. L-15 medium, adding different concentrations of FBS the taurine, Ca 2 , orthogonal experiment results showed that, 5? S beneficial to the explant cells move out, the higher the serum concentration cells move out of the worse effects. 5? S with the concentration of taurine combination can significantly prolong cell survival time of 1-2 weeks; high concentration of taurine (50 mM) of a more significant effect, cell survival extension 10 to the lower concentration of taurine (5mM, 20mM) group - the 30d about cardiomyocytes can maintain the normal growth of the state more than 60d. 20? Combination of S with various concentrations of taurine in the experimental group, the majority of cells survived for about two weeks, some of the myocardial cells can survive for a month or so, and these cells are more performance for the increase in size, contains a large number of particles in the cytoplasm; in the experimental group of the class combination of high concentrations of taurine (50 mM), the part of the cell with a higher succinic acid dehydrogenase activity, observed suspected portion telophase cells, and developmental phenomenon of early cell proliferation, but as the culture cell differentiation, gradually final vacuoles and degradation, culture Total maintain 41d. This is the first use of lentiviral vector-mediated attempt to the C. farreri SV40LT import primary cultured cardiomyocytes 3d, about 10d after infection was not observed significant cell proliferation. SV40LT transduction the C. farreri myocardial tissue in primary cultured cells in a variety of experimental conditions still need to further optimize.
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