Primary culture and histological characterization of phagocytic cells from rainbow trout Oncorhynchus mykiss

In this study the pronephros of rainbow trout Oncorhynchus mykiss was explanted and cultured. The morphology of cultured cells suggested they were leukocyte derived. To confirm this, cells were incubated with non-opsonized fluorescent beads and human apoptotic polymorphonuclear leukocytes (PMN) to establish that they were capable of phagocytosis and that they became activated following exposure to apoptotic bodies. The cells were studied using light microscopy, transmission electron microscopy and a live cell observer system. Cells in culture were also stained with phalloidin to identify actin reorganization following activation. Cultured cells ingested the inert beads and apoptotic PMNs and c. 80% of cells became activated following exposure to the apoptotic PMN, evidenced by enhanced filopodial extensions. This methodology may play a role in future studies, in particular, the competence of macrophages following exposure to radiation can now be tested.

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