Establishment and Characterization of a New Cell Line from the Kidney of Spotted Halibut Verasper variegates  

作　　者：WANG Xianli CHEN Songlin SHA Zhenxia FAN Tingjun WANG Na 

机构地区：[1]College of Marine Life Sciences, Ocean University of China, Qingdao 266003, P. R. China [2]Key Lab For Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, P. R. China

出　　处：《Journal of Ocean University of China》2010年第2期162-168,共7页中国海洋大学学报（英文版）

基　　金：supported by grants from State 863High-Technology Rand Project of China(2006AA09Z406,2006AA10A401);Taishan Scholar Project of Shan-dong Province

摘　　要：A cell line, SHK, was derived from the kidney of spotted halibut Verasper variegates. The cell line was subcultured more than 40 passages in minimum essential medium （MEM） supplemented with fetal bovine serum （FBS） and 10 ng ml4 basic fibroblast growth factor （bFGF）. Cell morphology from primary culture and subculture was observed continuously by microscopy. The SHK cell line consisted predominantly of fibroblast-like cells. The cell line was able to grow between 20℃ and 30℃ with the optimum growth at 24℃ and with a reduced growth between 12℃ and 20℃. The growth rate of the cells increased as the proportion of FBS increased from 10% to 20% at 28℃ with optimum growth at the concentration of 20%. The doubling time of the cells was determined to be 44.8 h. Chromosome analysis revealed that 52% of the SHK cells maintained a normal diploid chromosome number （2n=46）. The cells were successfully transfected with green fluorescent protein （GFP） reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. The cells were infected by lymphosystis disease virus （LCDV） and found to be susceptible to the virus in cytopathic effect （CPE） observation. The infection was confirmed by PCR and electron microscopy experiments, which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subculturedmore than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1basic fi-broblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.TheSHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃and 30℃with the optimum growth at 24℃and with a reduced growth between 12℃and 20℃.The growth rate of the cells increased as the proportion ofFBS increased from 10%to 20%at 28℃with optimum growth at the concentration of 20%.The doubling time of the cells was de-termined to be 44.8 h.Chromosome analysis revealed that 52%of the SHK cells maintained a normal diploid chromosome number(2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFPgene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis diseasevirus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCRand electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.

关 键 词：cell line spotted halibut Verasper variegates SHK cell line TRANSFECTION LCDV 

分 类 号：Q813[生物学—生物工程] Q959.4