文献类型：期刊文献

中文题名：四指马鲅稳传心脏细胞系的建立与应用

英文题名：Establishment and applications of a stable diploid heart cell line(EtH)from Eleutheronema tetradactylum

作者：李育浩[1,2];陈杨慧[1,2];张洪瑞[1,2];黄瑜[1,2];简纪常[1,2];王蓓[1,2];王忠良[1,2,3,4]

机构：[1]广东海洋大学水产学院,广东湛江524088;[2]广东海洋大学,广东省水产动物病害防控与健康养殖重点实验室,广东湛江524088;[3]广东省农业技术推广中心,广东广州510500;[4]广东海洋大学,广东省海洋渔业技术创新中心,广东湛江524088

年份：2026

卷号：50

期号：7

起止页码：57

中文期刊名：水产学报

外文期刊名：Journal of Fisheries of China

收录：北大核心2023、、北大核心

基金：广东省现代化海洋牧场适养品种核心技术攻关项目(2024-MRB-00-001);广东省普通高校重点领域专项(2023ZDZX4011)。

语种：中文

中文关键词：四指马鲅;心脏细胞系;重金属毒性;转染;细菌敏感性

外文关键词：Eleutheronema tetradactylum;heart-derived cell line;heavy metal toxicity;transfection;bacterial susceptibility

中文摘要：【目的】建立四指马鲅细胞系旨在提供非模式经济鱼类细胞模型的研究体系,为后续深入开展基因功能解析与种质改良研究开辟新路径。【方法】以四指马鲅心脏组织为研究对象,采用蛋白酶消化法进行原代培养,并对该细胞系进行细胞系鉴定与基础应用。使用不同培养条件对细胞系进行生长条件筛选,冻存细胞后检测复苏率,并对细胞系进行核型分析与细胞来源鉴定。使用Lipo3000试剂对细胞系进行转染,分别配制3种含不同浓度重金属的培养基对细胞系进行重金属敏感性测试,并研究细胞系对两种不同的细菌的敏感性。【结果】目前该细胞系已稳定传至70代(命名为EtH),该细胞系为正常二倍体(2N=26),冻存复苏后存活率达81.3%以上;最适生长条件为L-15培养基、10%胎牛血清(FBS)、28℃培养温度。细胞转染效率约16%,对锰、铬、铜3种重金属表现出剂量依赖性毒性敏感,对无乳链球菌和诺卡氏菌均敏感。【结论】EtH细胞系具有相对稳定的遗传性,冻存后仍保持较高的存活率,EtH细胞系可作为重复使用的研究工具。该细胞系对不同种类重金属与细菌展现出特异敏感性,在环境污染物监测和毒理学领域以及四指马鲅等鱼类病害学相关实验中具备良好应用潜力。通过脂质体转染法成功导入pEGFP-N1质粒,更精准检测出EGFP基因约16%的表达率。因此,该细胞系可以作为研究外源性基因的体外研究工具。相较于当前鱼类细胞系研究中“偏向模式物种”的现状,本研究填补了四指马鲅的“专属”细胞系的空白,丰富了非模式经济鱼类细胞模型的研究体系,为后续基因功能解析与种质改良提供了可落地的研究路径。

外文摘要：The four-finger threadfin(Eleutheronema tetradactylum)is a marine fish species of high economic value.Although artificial breeding and selection have been achieved in China,challenges such as high stress susceptibility and diseases persist in aquaculture.This study aimed to establish a cell line from this species to provide a research platform for non-model economic fish,paving a new path for in-depth gene function analysis and genetic improvement.Heart tissue from E.tetradactylum was used for primary culture via protease digestion.The established cell line was then systematically characterized.Optimal growth conditions were determined by testing different culture parameters.Cells were cryopreserved to assess recovery rate,and karyotype and species origin analyses were performed.The cell line was transfected using Lipofectamine 3000.Sensitivity tests were conducted using media containing three heavy metals at varying concentrations,as well as against two bacterial species.A stable cell line,designated EtH,has been subcultured to passage 70.It exhibits a normal diploid karyotype(2N=26)and a post-thaw viability exceeding 81.3%.The optimal growth conditions were identified as Leibovitz's L-15 medium supplemented with 10%fetal bovine serum at 28°C.The transfection efficiency for a pEGFP-N1 plasmid was approximately 16%.The EtH cells showed dose-dependent toxic sensitivity to manganese,chromium,and copper,and were susceptible to both Streptococcus agalactiae and Nocardia sp..The EtH cell line demonstrates stable genetic characteristics and high post-thaw viability,making it a reliable and reusable research tool.Its specific sensitivity to various heavy metals and bacteria indicates strong application potential in environmental pollutant monitoring,toxicology,and fish disease research for E.tetradactylum and related species.Successful transfection confirms its utility as an in vitro tool for exogenous gene expression studies.Addressing the current research bias towards model species,this study fills the gap of a"dedicated"cell line for E.tetradactylum,enriches the research system for non-model economic fish cell models,and provides a practical pathway for future gene function analysis and germplasm improvement.