水产学报2018，Vol.42Issue(3)：376-387,12.DOI:10.11964/jfc.20170410798
永生化尼罗罗非鱼巨噬细胞系的建立及鉴定
Establishment and identification of immortalized macrophage cell line of tilapia(Oreochromis niloticus)
摘要
Abstract
To obtain a continuous monoclonal Oreochromis niloticus macrophage cell line,macrophages were isol-ated and purified from peritoneal cavity of O.niloticus.The O.niloticus macrophage cell line was established and EBV infection identification, electron microscopy, telomerase activity assay, carcinogenicity evaluation, karyo-type analysis, and molecular biology identification were then carried out. The results showed that EBV had been integrated into O.niloticus peritoneal macrophages and stably expressed.After 30 generations of stable passage, the cell line still maintained a good proliferative state. The cell line surface was not smooth and had obvious blunt round protrusion and slender pseudopodia, which was the typical morphology performance of the macrophages. Telomerase activity of this cell line was significantly higher than that of untreated macrophages (P<0.05), but the difference was not significant with HeLa cells, and the cell line was not carcinogenic, which indicated that immor-talized cell lines were successfully constructed. Karyotype analysis showed that the cell line had 44 chromosomes, and the karyotype formula was 2 n=2 x=44=4 sm+17 st+1 t.PCR detection revealed that CD33,and CD205 tran-scripts were found in this cell line,all of which were markers of monocytes/macrophages.18S rRNA detection showed that the cell line was from O.niloticus macrophages.Immortalized tilapia macrophage cell lines were suc-cessfully established. The establishment of this cell line laid the foundation for the further study of the immunolo-gical function of peritoneal macrophages in O.niloticus and also provided a tool for studying the mechanism of high protection rate of O.niloticus Streptococcus HSP70-peptide vaccine.关键词
尼罗罗非鱼/巨噬细胞/永生化/细胞系Key words
Oreochromis niloticus/macrophages/immortalization/cell line分类
农业科技引用本文复制引用
张永德,潘传燕,余艳玲,冯鹏霏,罗洪林..永生化尼罗罗非鱼巨噬细胞系的建立及鉴定[J].水产学报,2018,42(3):376-387,12.基金项目
国家自然科学基金(31372553) （31372553）
广西自然科学基金(2015GXNSFAA139068) （2015GXNSFAA139068）
广西科技计划基地和人才专项重点实验室建设项目(16-380-38)National Natural Science Foundation of China(31372553) （16-380-38）
Natural Science Foundation of Guangxi Province(2015GXNSFAA139068) （2015GXNSFAA139068）
Key Laboratory Construction Project of Guangxi Science and Tech-nology Base and Talent(16-380-38) （16-380-38）
