| 大黄鱼头肾巨噬细胞体外模型的构建 |
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| 引用本文： | 李庆飞,艾庆辉,麦康森,郑岳夫,徐玮,张文兵. 大黄鱼头肾巨噬细胞体外模型的构建[J]. 水生生物学报, 2013, 37(5): 885-891. DOI: 10.7541/2013.113 |
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| 作者姓名： | 李庆飞 艾庆辉 麦康森 郑岳夫 徐玮 张文兵 |
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| 作者单位： | 1. 中国海洋大学海水养殖教育部重点实验室,山东 青岛,266003
2. 象山县海洋渔业局,浙江 象山,315700 |
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| 摘 要： | 通过优化Percoll密度梯度离心技术,从大黄鱼 (Larmichthys crocea) 头肾组织中分离纯化巨噬细胞,并优化细胞培养条件。对所分离细胞进行形态学分析、普通光镜和电镜超微结构观察以及细胞功能验证实验。实验结果表明:细胞单层置于22℃无CO2恒温培养箱中,于L15培养基中培养5d后仍保持较高的活力。普通光镜和电镜观察到大黄鱼头肾巨噬细胞具有伪足发生和较强的吞噬能力、胞浆中富含线粒体以及吞噬小泡。巨噬细胞与不同浓度(0、0.1、1、20 μg/mL)的脂多糖(LPS,Escherichia coli 055:B5)分别孵育3h和24h。施加LPS刺激后,细胞呼吸暴发活性随LPS浓度以及孵育时间变化而变化。孵育3h后,0.1 μg/mL LPS协同PMA可显著诱导巨噬细胞活性氧中间体(ROI)的产生。孵育24h后,所有处理组LPS协同PMA显著抑制细胞ROI的生成(P 0.05)。未添加PMA时, 经1 μg/mL LPS孵育3h和24h后的巨噬细胞细胞ROI的生成量显著降低(P 0.05), 其他LPS处理组间细胞ROI的生成量无显著差异。
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| 关 键 词： | 大黄鱼 巨噬细胞 吞噬 呼吸暴发 离体 |
DEVELOPMENT OF A SHORT-TERM IN VITRO MODEL OF MACROPHAGES FROM LARGE YELLOW CROAKER, LARMICHTHYS CROCEA (RICHARDSON, 1846) |
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| LI Qing-Fei , AI Qing-Hui , MAI Kang-Sen , ZHENG Yue-Fu , XU Wei , ZHANG Wen-Bing. DEVELOPMENT OF A SHORT-TERM IN VITRO MODEL OF MACROPHAGES FROM LARGE YELLOW CROAKER, LARMICHTHYS CROCEA (RICHARDSON, 1846)[J]. Acta Hydrobiologica Sinica, 2013, 37(5): 885-891. DOI: 10.7541/2013.113 |
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| Authors: | LI Qing-Fei AI Qing-Hui MAI Kang-Sen ZHENG Yue-Fu XU Wei ZHANG Wen-Bing |
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| Abstract: | Head kidney macrophages from large yellow croaker were isolated by density gradient sedimentation and characterized under light microscope and electronic microscope. Cells were maintained in Leibowitz L-15 medium for 5 days at an optimal temperature of 22℃. The typical macrophages, rich in phagosomes and mitochondria, exhibited strong adhesion and plasticity as well as high phagocytic capacity of engulfing yeasts. Macrophages were incubated with lipopolysaccharide (LPS) from Escherichia coli 055:B5 (0, 0.1, 1, 20 μg/mL) for 3h and 24h, respectively. After the exposure, reactive oxygen intermediates (ROI) production was determined. Only 3h incubation with 0.1 μg/mL LPS could significantly enhance ROI production of PMA-triggered macrophages compared to control cells (LPS = 0 μg/mL), but after 24h incubation, ROI production was significantly decreased by all levels of LPS (P 0.05). At the absence of PMA, significant decrease in ROI production was found in the treatment with 1 μg/mL LPS, while no obvious effects were observed in treatments with other levels of LPS. |
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| Keywords: | Large yellow croaker (Larmichthys crocea) Macrophages Phagocytosis Respiratory burst activity In vitro |
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