| 摘要: |
| 草鱼(Ctenopharyngodon idellus)性腺细胞系(GCO)是中国科学院水生生物研究所在20世纪70年代开展草鱼出血病研究时建立的一株细胞系，迄今已传至300多代，在中国鱼类病毒学研究领域发挥了重要作用。本研究采用形态学观察、细胞生长曲线测定、细胞周期测定、细胞核型分析、细胞凋亡检测、电镜观察等方法，对GCO的生长特性及鲤春病毒血症病毒(SVCV)在该细胞中的增殖特性等进行了研究。结果显示，GCO细胞的最适生长温度为25℃，在M199和MEM等细胞培养液中均能较好地生长，培养液中最适的胎牛血清浓度为10%。测定了GCO细胞系对SVCV病毒的敏感性，发现与鲤(Cyprinus carpio)上皮瘤细胞系(EPC)、肥头鲤(Pimephales promelas)细胞 系(FHM)、大鳞大麻哈鱼(Oncorhynchus tshawytscha)胚胎细胞系(CHSE-214)等世界动物卫生组 织(OIE)推荐和各检测实验室常用的鱼类细胞系相比，GCO细胞系对SVCV表现出非常高的敏感性。生长曲线、电镜观察和凋亡实验显示，SVCV能引起GCO细胞系出现明显而稳定的细胞病变，引起GCO细胞系出现凋亡，并在细胞质中大量增殖。结果表明，GCO细胞系适用于SVCV病毒的分离、检测以及病毒致病性的有关研究。GCO细胞的适宜生长温度为15–28℃，这一特点将使它可以广泛地用于多种水生动物病毒的分离。 |
| 关键词: 草鱼性腺细胞系 生物学特性 易感性 鲤春病毒血症病毒 |
| DOI：10.11758/yykxjz.20151119001 |
| 分类号: |
| 基金项目:国家质检总局科技计划项目(2014IK236)、国家质检总局公益性行业科研专项(201210055；20120214)和深圳检验检疫局科技计划项目(SZ2014202)共同资助 |
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| Characterization of Grass Carp (Ctenopharyngodon idellus) Ovary Cell Line and Its Susceptibility to Spring Viremia of Carp Virus |
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WANG Jinjin, LIU Ying, YU Li, JIA Peng, CHEN Bing, SHI Xiujie, ZHENG Xiaocong, LAN Wensheng, HE Junqiang, LIU Hong
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Animal & Plant Inspection and Quarantine Technology Center, Shenzhen Entry-Exit Inspection and Quarantine Bureau of P.R. China, Shenzhen 518045
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| Abstract: |
| Grass carp ovary cell lines (GCO) was used in the study of Grass Carp Hemorrhage and has been established by Chinese Academy of Sciences Institute of Aquatic Animal in 1970s. GCO has been cultured for more than 300 generations and plays an important role in fish virology research in China. In this study, we performed morphological observation, drawing of the growth curves, cell cycle analysis, chromosomal analysis, cytopathic effect assay (CPE), TCID50 assay, cell apoptosis detection, and electron microscopic observation to demonstrate the characteristics of GCO and the cell proliferation of spring viremia of carp virus (SVCV). It was observed that GCO cell line mainly consisted of epithelial-like cells. GCO grew at temperatures between 15℃ and 28℃, and the optimum was 25℃. The growth rate of GCO cells increased proportionally with the foetal bovine serum (FBS) concentration (5% to 20%), and the optimum concentration of FBS was 10%. There were 48 chromosomes in GCO cells. Four known fish cell lines were tested to determine the susceptibility of SVCV, and the results demonstrated that GCO was susceptible to SVCV. Moreover, GCO displayed higher sensitivity to SVCV and increased viral production than the control cell line, epithelioma papulosum cyprini (EPC), fathead minnow (FHM), and chinook salmon embryo (CHSE-214) cell lines. These results suggested that GCO cell line could be a useful diagnostic tool in SVCV isolation and diagnosis, and could serve as a suitable biological system in the pathogenicity study of SVCV. One advantage of this cell line is that it grows in a wide temperature range (15℃ to 28℃), and thus can be used in isolating both warm-water and cold-water fish viruses. These characteristics of the GCO make it a potential tool in the future study of other fish viruses. |
| Key words: Grass carp ovary cell lines (GCO) Biological characteristics Susceptibility Spring viraemia of carp virus (SVCV) |
