鳗鲡肾脏细胞系的建立及其对鳗鲡疱疹病毒的敏感性

THE ESTABLISHED KIDNEY CELL LINE FROM EUROPEAN EEL ANGUILLA ANGUILLA AND ITS SUSCEPTIBILITY TO ANGUILLID HERPESVIRUS

为开展鳗鲡病毒性疾病流行病学调查和病原生物学研究,进行鳗鲡病毒的分离、培养和鉴定,研究采用组织块培养法,以欧洲鳗鲡肾脏组织为材料,建立了欧洲鳗鲡肾脏细胞系(European eel kidney cell line,EEK),EEK形态呈类纤维状,经过约12个月的培养,已成功传至38代。通过对其培养液、血清浓度和培养温度等条件进行优化,发现DMEM/F12、L15培养液均适合其正常生长和增殖,而在MEM培养液中无法正常生长;在5%—15%FBS(Fetal Bovine Serum)浓度范围内,其生长速度随FBS浓度的升高而增快,当FBS浓度高于20%或过低于5%时,其生长速率有所减慢;在22—27℃时生长良好,但当温度低于17℃和高于32℃时细胞生长速度明显减慢。对EEK细胞进行鳗鲡疱疹病毒(Anguillid herpesvirus,AnHV)敏感性实验,结果表明该细胞系对鳗鲡疱疹病毒敏感,可产生明显的细胞病变。EEK的建立丰富了鱼类细胞系的种类,为鳗鲡病毒性疾病诊断、病毒性病原学研究和病毒疫苗研制提供了重要实验材料。

To investigate the epidemiology and etiology of eel viral diseases,isolate,cultivate and identify the eel virus,a new cell line(EEK)was established by tissue explant method from European eel Anguilla anguilla.EEK cells were fibroblast-like cells and were maintained and subcultured 38 generations over a 12-months period.The optimization of medium,serum concentration and temperature was conducted.EEK cells can grow and proliferate normally in DMEM/F12 and L15,but can not proliferate in MEM.The growth rate enhanced with the increased Fetal Bovine Serum(FBS)in the range of 5%—15%,and it decreased with more than 20%FBS and less than 5%FBS.The cells grew well in a temperature range of 22—27℃,but the growth rate reduced at the temperature below 17℃or above 32℃.The virus sensitivity test revealed that the cell line was susceptible to the infection of Anguillid herpesvirusv(An-HV)and has obvious cytopathic effect.The establishment of a kidney cell line from european eels has increased the variety of fish cell lines,providing important experimental material for the diagnosis of eel viral diseases,the study of viral etiology and the development of virus vaccine.