罗非鱼腹腔巨噬细胞分离与培养

摘要

为获得罗非鱼腹腔巨噬细胞体外分离培养条件和培养形态特征,试验在不同饲养温度和时间点注射角鲨烯的奥尼罗非鱼进行腹腔巨噬细胞分离；通过培养基血清筛选和瑞氏染色对分离的巨噬细胞进行了体外培养及其形态观察.结果表明,19～25℃饲养温度有利于腹腔巨噬细胞分离,大于28℃对分离效果有明显影响；注射角鲨烯后48～72 h是分离腹腔巨噬细胞最佳时期；体外培养显示自体血清有利于罗非鱼腹腔巨噬细胞的存活与生长,胎牛血清不适于罗非鱼腹腔巨噬细胞培养.同时,罗非鱼腹腔巨噬细胞体外培养具有哺乳动物巨噬细胞相似特征,形态不规则、核浆比值低、贴壁生长、可形成多核巨大细胞.为下一步研究罗非鱼腹腔巨噬细胞免疫学功能奠定了基础.%In order to study factors that influence the separation and culture of tilapia peritoneal macrophages, and to recognition the characteristic of tilapia peritoneal macrophages . In the study, We separated macrophages from peritoneal cavity of Tialpia( Oreochromis niloticus ♀× Oreochromis aureus ♂ ) after intraperitoneal injected with squa lene in the condition of difference growth temperature; The cell characteristic was observed by screening cell medi um and Wright's staining. The results show the macrophages were easily obtained in the condition of 19 ~25t , Separation result was obviously affected when water temperature was aboved 28 ℃. ;It was a optimization period for separation between 48-72 h after injected squalene ; Serum of tilapia play an important role in the tilapia perito neal macrophages invitro cultrue and fetal bovine serum was not fit;The characteristic of tilapia peritoneal macro phage was similar to mammalian macrophage, irregularly shaped with a low nucleus to cytoplasm ratio, adherent to plastic and glass,able to form multi-nucleated giant. The work laid a foundation for the further research of immuno logical function about tilapia peritoneal macrophage.