水产学报2026，Vol.50Issue(7)：57-70,14.DOI:10.11964/jfc.20251115205
四指马鲅稳传心脏细胞系的建立与应用
Establishment and applications of a stable diploid heart cell line(EtH)from Eleutheronema tetradactylum
摘要
Abstract
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 eco-nomic 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 spe-cies.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 supplemen-ted with 10%fetal bovine serum at 28℃.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 Strepto-coccus 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.Address-ing 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 func-tion analysis and germplasm improvement.关键词
四指马鲅/心脏细胞系/重金属毒性/转染/细菌敏感性Key words
Eleutheronema tetradactylum/heart-derived cell line/heavy metal toxicity/transfection/bacterial susceptibility分类
生物科学引用本文复制引用
李育浩,陈杨慧,张洪瑞,黄瑜,简纪常,王蓓,王忠良..四指马鲅稳传心脏细胞系的建立与应用[J].水产学报,2026,50(7):57-70,14.基金项目
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) （2024-MRB-00-001）
广东省普通高校重点领域专项(2023ZDZX4011) Research on Breeding Technology of Candidate Species for Guangdong Modern Marine Ranching(2024-MRB-00-001) （2023ZDZX4011）
Guangdong Province Ordinary Colleges and Universities Key Field Special Project(2023ZDZX4011) （2023ZDZX4011）
