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斑点狼鱼(Anarhichas minor Olafsen)巨噬细胞的分离与鉴定

Isolation and characterisation of spotted wolffish (Anarhichas minor Olafsen) macrophages.

作者信息

Norum M, Bøgwald J, Dalmo R A

机构信息

Department of Marine Biotechnology, Institute of Marine Biotechnology, Norwegian College of Fishery Science, University of Tromsø, N 9037 Tromsø, Norway.

出版信息

Fish Shellfish Immunol. 2005 May;18(5):381-91. doi: 10.1016/j.fsi.2004.10.002.

Abstract

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.

摘要

非特异性机制在所有多细胞动物抵御病原微生物的过程中起着重要作用。巨噬细胞和粒细胞在这方面发挥着核心作用。因此,开发获取和培养巨噬细胞并评估其在斑点狼鱼(一种当前水产养殖业感兴趣的冷水物种)中功能的方法是很有必要的。通过使用Percoll密度沉降法从斑点狼鱼(Anarhichas minor Olafsen)的肾脏中分离出巨噬细胞。这些细胞具有高度吞噬能力,并通过透射电子显微镜和扫描电子显微镜评估具有典型的巨噬细胞形态。通过电子显微镜分析,从Percoll密度界面收集的巨噬细胞大小为5 - 9微米。接种24小时后测量,当细胞在添加合成血清替代品(SSR - 2)的情况下于13℃保存时,体外活力最高(87.1%)。与对照细胞相比,通过硝基蓝四唑还原法测量,添加细菌脂多糖(LPS)24小时后,1日龄细胞未被显著激活。这些细胞在补体成分C3的合成和含量方面呈阴性。

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