Isolation and characterisation of spotted wolffish (Anarhichas minor Olafsen) macrophages.
Non-specific mechanisms are important in the defence of all multicellular animals against pathogenic microorganisms. Macrophages and granulocytes play a central role in this respect. It is thus pertinent to develop methods for obtaining and cultivation of macrophages and assessing their functions in the spotted wolffish, a cold water species of current interest for the aquaculture industry. Kidney macrophages from the spotted wolffish (Anarhichas minor Olafsen) were isolated by density sedimentation using Percoll. The cells were highly phagocytic and possessed typical macrophage morphology evaluated by transmission and scanning electron microscopy. Using electron microscopic analysis, the size of the macrophages, collected from the Percoll density interface, was 5-9 microm. The viability in vitro was highest (87.1%) when the cells were kept at 13 degrees C with the addition of synthetic serum replacement (SSR-2) when measured 24 h after seeding. One day old cells were not significantly activated by addition of bacterial lipopolysaccharide (LPS) for 24 h when measured by reduction of nitroblue tetrazolium compared to control cells. The cells were negative in respect to synthesis and contents of complement component C3.
Norum
M
M
Department of Marine Biotechnology, Institute of Marine Biotechnology, Norwegian College of Fishery Science, University of Tromsø, N 9037 Tromsø, Norway.
Bøgwald
J
J
Dalmo
R A
RA
D000818:Animals; D017931:DNA Primers; D007150:Immunohistochemistry; D007668:Kidney / Q000276:immunology; D008070:Lipopolysaccharides; D008264:Macrophages / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009580:Nitroblue Tetrazolium; D010473:Perciformes / Q000235:genetics / Q000276:immunology*; D010587:Phagocytosis / Q000502:physiology; D011205:Povidone; D012333:RNA, Messenger / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012822:Silicon Dioxide
D017931:DNA Primers; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D009580:Nitroblue Tetrazolium; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone