齐口裂腹鱼鳃丝细胞系的建立、鉴定及免疫研究  

Establishment,identification,and immunological study of gill filament cell line of Schizothorax prenanti

作　　者：巩志宏 王娜[2] 刘娟娟 胡国斌[1] 陈松林[2] GONG Zhihong;WANG Na;LIU Juanjuan;HU Guobin;CHEN Songlin(College of Marine Life,Ocean University of China,Qingdao 266071,China;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;Research Institute of Chinese Sturgeon,China Three Gorges Corporation,Yichang 443100,China)

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266071 [2]中国水产科学研究院黄海水产研究所,山东青岛266071 [3]中国长江三峡集团有限公司中华鲟研究所,湖北宜昌443100

出　　处：《中国水产科学》2022年第10期1407-1416,共10页Journal of Fishery Sciences of China

基　　金：国家海洋水产种质资源库项目;中国水产科学研究院创新团队专项(2020TD20);山东省泰山学者攀登计划项目;三峡工程鱼类资源保护湖北省重点实验室开放课题项目(191164740001)

摘　　要：齐口裂腹鱼(Schizothorax prenanti)是长江上游水域的珍稀鱼类,近年来由于环境变化和疾病爆发野外齐口裂腹鱼数量急剧减少,现已被列入长江上游二级急切保护的特有鱼类,因此,对齐口裂腹鱼的保护刻不容缓。建立齐口裂腹鱼细胞系是保护其种质资源的有效手段,也可以在不伤害现有鱼群的条件下进行多种齐口裂腹鱼相关生物学研究。本研究建立了首个来源于齐口裂腹鱼的细胞系,SPG细胞系。原代细胞分离自齐口裂腹鱼鳃丝组织,呈均一的上皮状,使用含15%血清的L-15培养,在15个月时间里成功传至55代。线粒体COI基因鉴定,证明该细胞来源于齐口裂腹鱼,核型检测细胞染色体数目为2n=96。在液氮中保存12个月的细胞,复苏后能保持75%以上活力。EGFP-N3质粒转染SPG细胞后观察到明显的绿色荧光蛋白表达。病毒类似物poly(I:C)和大肠杆菌脂多糖LPS可引起细胞IL-1β、IL-8、TNFa和TLR22等免疫相关基因表达量升高。表明本研究建立的齐口裂腹鱼鳃丝细胞系可用于免疫学研究。此外,此细胞系还将在种质保存,外源蛋白表达和齐口裂腹鱼体外生物学研究中发挥重要作用。Schizothorax prenanti is a rare fish in the upper reaches of the Yangtze River and used to be an important economic fish in its production area.However,in recent years,due to diseases and environmental changes,the number of Schizothorax prenanti in the wild has decreased sharply.Thus,there is an urgent need to protect this fish species.The establishment of Schizothorax cell lines is a cost-effective method to protect its germplasm resources.Since the establishment of the fish cell line RTG-2 in 1962,research on the establishment of fish cell lines has developed rapidly.A total of 783 cell lines have been established from various fish tissues,including embryos.However,thus far,reports on the establishment of cell lines from Schizothorax prenanti are lacking.The establishment of the Schizothorax prenanti cell line not only enables the protection of its germplasm resources but also facilitates research on its living environment toxicology,nutrition,immunology,and disease control without harming the existing population of this rare fish.Schizothorax prenanti can be infected with Streptococcus agalactiae in the natural environment,resulting in exophthalmos,surface hemorrhage,and neurological symptoms,as well as marked degeneration,necrosis,and inflammatory cell infiltration in various internal organs.Mucosal immunity is an important part of the immune system of this fish,wherein the gills,skin,and intestinal mucosa constitute the first line of defense against pathogenic microorganisms.Thus,the establishment of a stable growth of the gill filament cell line of Schizothorax prenanti will enable the study of immune mechanism and disease control of Schizothorax prenanti.The most commonly used media for fish cell culture are MEM and L-15,where L-15 is an amino acid-rich medium that does not require CO_(2)to maintain the pH of the medium.Among the 280 newly established fish cell lines in the world from 2010 to 2020,180 use L-15medium,accounting for 64%of the total.In this study,L-15 was used as the basis to prepare the Schi

关 键 词：齐口裂腹鱼 SPG 细胞系 免疫研究 

分 类 号：S917.4[农业科学—水产科学]