金鱼疱疹病毒敏感细胞的体外培养

In vitro culture of goldfish cell sensitive to goldfish herpes virus

采用组织块法和胰蛋白酶消化法对金鱼细胞进行体外培养和传代,研究了高倍稀释、培养液中的小牛血清浓度以及冷冻保存等因子对显示出较高的增殖能力的来源于鳍的细胞GFF的影响进行了试验;并对其进行金鱼疱疹病毒接种、病毒传代培养等试验和电子显微镜观察。结果不论是高倍稀释,还是在低浓度小牛血清培养液H MEM 5中培养,细胞均具有足够的增殖速率;而且冷冻保存在细胞在解冻后的培养仍然具有较好的增殖能力,细胞的存活率在85%以上,使此细胞的冷冻保存成为可能;同时,接种细胞出现了CPE且病毒传代培养的细胞内和培养液中均观察到病毒颗粒,表明GFF细胞可以支持病毒的增殖,为研究金鱼病毒奠定了基础。

By using the technique of tissue culture, we successfully obtained the cell from goldfish fin, skin, eye, heart, spleen, spermary, swim bladder and kidney. Among them, GFF(cell from goldfish fin)has the strongest proliferating ability. Tests of high times dilution, decreasing the percent of newborn calf serum and freezing conservation with GFF indicate that high times dilution cell still have the normal proliferating ability; GFF can be cultured in HMEM5; after defrosting, GFF still maintained high proliferating ability and the cell survival rate is 85%. At the same time, GFF was inoculated with herpes virus of goldfish, after CPE appeared, the passaged virus was observed by electron microscope. The results indicate that GFF cell can sustain the propagation of herpes virus of goldfish, and this established the basis of researches about goldfish virus.