Sue-Min Huang, C. Tu, S. Kuo, H. Kuo, C. Chou, Shao-Kuang Chang
{"title":"石斑鱼肾细胞系的建立、鉴定及其对石斑鱼虹膜病毒的敏感性","authors":"Sue-Min Huang, C. Tu, S. Kuo, H. Kuo, C. Chou, Shao-Kuang Chang","doi":"10.4172/2155-9910.1000192","DOIUrl":null,"url":null,"abstract":"A new continuous cell line, designated as GK-7, was developed from the kidney tissue of the marine grouper, Epinephelus coioides. The cell line grew well in Leibovitz’s L-15 medium supplemented with 15% fetal bovine serum at a range of temperatures from 20 to 32â, with optimal growth at 25â. Morphologically, the GK-7 cell line is spindleshaped epithelial-like cell comfirmed by immunophenotyping with cytokeratin antibody. Chromosome number analysis showed that GK-7 cells of the 50th and 150th cell passages had a modal diploid chromosome number of 48 and 66, respectively. Replication of GIV with the cell line showed that the maximum virus yield reached up 108.4 TCID50 mL-1 with the cells of the 50th passage. Electron micrographs showed abundant cytoplasmic icosahedral virions with a mean diameter of 200 nm in virus-infected cells. Negative staining of ultrathin sections of infected cells showed three-layered membrane enveloped mature viral particles with a diameter about 240 nm. Green fluorescent protein can be expressed in both cell lines at 48 hr after the cell lines were transfected with a green fluorescent reporter gene driven by a cytomegalovirus promoter. Our results showed that the GK-7 cell line provided valuable tools for the isolation and investigation of fish iridovirus and for vaccine production.","PeriodicalId":331621,"journal":{"name":"Journal of Marine Science: Research & Development","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2016-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"4","resultStr":"{\"title\":\"Establishment and Characterization of a Novel Kidney-cell Line fromOrange-spotted Grouper, Epinephelus coioides, and its Susceptibility toGrouper Iridovirus\",\"authors\":\"Sue-Min Huang, C. Tu, S. Kuo, H. Kuo, C. Chou, Shao-Kuang Chang\",\"doi\":\"10.4172/2155-9910.1000192\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"A new continuous cell line, designated as GK-7, was developed from the kidney tissue of the marine grouper, Epinephelus coioides. The cell line grew well in Leibovitz’s L-15 medium supplemented with 15% fetal bovine serum at a range of temperatures from 20 to 32â, with optimal growth at 25â. Morphologically, the GK-7 cell line is spindleshaped epithelial-like cell comfirmed by immunophenotyping with cytokeratin antibody. Chromosome number analysis showed that GK-7 cells of the 50th and 150th cell passages had a modal diploid chromosome number of 48 and 66, respectively. Replication of GIV with the cell line showed that the maximum virus yield reached up 108.4 TCID50 mL-1 with the cells of the 50th passage. Electron micrographs showed abundant cytoplasmic icosahedral virions with a mean diameter of 200 nm in virus-infected cells. Negative staining of ultrathin sections of infected cells showed three-layered membrane enveloped mature viral particles with a diameter about 240 nm. Green fluorescent protein can be expressed in both cell lines at 48 hr after the cell lines were transfected with a green fluorescent reporter gene driven by a cytomegalovirus promoter. Our results showed that the GK-7 cell line provided valuable tools for the isolation and investigation of fish iridovirus and for vaccine production.\",\"PeriodicalId\":331621,\"journal\":{\"name\":\"Journal of Marine Science: Research & Development\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-05-31\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"4\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Marine Science: Research & Development\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.4172/2155-9910.1000192\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Marine Science: Research & Development","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.4172/2155-9910.1000192","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Establishment and Characterization of a Novel Kidney-cell Line fromOrange-spotted Grouper, Epinephelus coioides, and its Susceptibility toGrouper Iridovirus
A new continuous cell line, designated as GK-7, was developed from the kidney tissue of the marine grouper, Epinephelus coioides. The cell line grew well in Leibovitz’s L-15 medium supplemented with 15% fetal bovine serum at a range of temperatures from 20 to 32â, with optimal growth at 25â. Morphologically, the GK-7 cell line is spindleshaped epithelial-like cell comfirmed by immunophenotyping with cytokeratin antibody. Chromosome number analysis showed that GK-7 cells of the 50th and 150th cell passages had a modal diploid chromosome number of 48 and 66, respectively. Replication of GIV with the cell line showed that the maximum virus yield reached up 108.4 TCID50 mL-1 with the cells of the 50th passage. Electron micrographs showed abundant cytoplasmic icosahedral virions with a mean diameter of 200 nm in virus-infected cells. Negative staining of ultrathin sections of infected cells showed three-layered membrane enveloped mature viral particles with a diameter about 240 nm. Green fluorescent protein can be expressed in both cell lines at 48 hr after the cell lines were transfected with a green fluorescent reporter gene driven by a cytomegalovirus promoter. Our results showed that the GK-7 cell line provided valuable tools for the isolation and investigation of fish iridovirus and for vaccine production.