DAO 125:217-226 (2017)  -  DOI: https://doi.org/10.3354/dao03145

ABSTRACT: Primary cultured cells can be a useful tool in studies on physiology, virology, and toxicology. Hemocytes play an important role in animal rapid response to pathogen invasion. In this study, an appropriate medium for primary culture of hemocytes of the bivalve Chlamys farreri was developed by adding 5% fetal bovine serum and 1% C. farreri serum to Leibovitz L-15 medium. These primary cultured hemocytes were maintained for more than 40 d in vitro and were classified into 3 types: (1) granulocytes containing numerous granules in the cytoplasm, (2) hyalinocytes with no or few granules, (3) a small percentage of macrophage-like cells. Furthermore, the primary cultured hemocytes were observed to be sensitive to bacterial and viral challenges. These hemocytes could phagocytose the bacterium Vibrio anguillarum, and presented cytopathic effects on the extracellular products (ECPs) of V. anguillarum; the mRNA level of QM, which plays an important role in immune response, also significantly increased 12 h after infection. When these hemocytes were challenged with ostreid herpesvirus 1 (OsHV-1), virus particles and empty capsids in the cells infected for 48 h were observed by transmission electron microscopy, and the QM mRNA level increased significantly at 12 h and 24 h following OsHV-1 challenge. This primary culture system is available for C. farreri hemocytes which can be used in the future to study host-pathogen interactions.

KEY WORDS: Chlamys farreri · Cell culture · Hemocyte · Vibrio anguillarum · Ostreid herpesvirus 1