Two cell lines were established from silver crucian carp and goldfish brain tissue and used as the biological tool for monitoring viral diseases. Characterization including optimal growth kinetics study, karyotyping, and mitochondrial ribosomal RNA (rRNA) genotyping were performed. The primary cultures of these cells were generated by the explant technique using the medium 199 supplemented with 20 % fetal bovine serum and epidermal/fibroblast growth factors. The cells grew over the range of 15 to 30°C, while the optimal temperature for culture was 30°C. The cell lines were maintained in vitro and could be subcultured over 40 times. Following cryopreservation in liquid nitrogen, thawed cells exhibit viability of > 90 % after a 13-mo period of storage. The chromosome count of two cell lines were determined to be 154 and 110, respectively, which agreed well with triploid crucian carp brain cells and diploid goldfish brain cells. Polymerase chain reaction amplification and sequence analysis indicated 100 % and 94% match with known crucian carp mitochondrial DNA sequences. Cytopathic effect was continuously observed in both cell lines over 10 passages after inoculation with tissue homogenates of sick or died goldfish from cyprinid herpesvirus 2 (CyHV-2) outbreaks. These newly established cell lines could be a diagnostic tool for viral diseases in fish species.

中文翻译：

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金鱼和silver鱼的两种脑细胞系的发育以及对赛普拉斯疱疹病毒2的病毒敏感性。

从silver鱼和金鱼的脑组织中建立了两个细胞系，并将其用作监测病毒性疾病的生物学工具。进行了表征，包括最佳生长动力学研究，核型分析和线粒体核糖体RNA（rRNA）基因分型。这些细胞的原代培养物是通过外植技术使用添加有20％胎牛血清和表皮/成纤维细胞生长因子的培养基199生成的。细胞在15至30°C的范围内生长，而培养的最佳温度为30°C。细胞系保持体外，可以传代培养40次以上。在液氮中冷冻保存后，解冻的细胞在保存13个月后显示出大于90％的活力。确定两个细胞系的染色体数分别为154和110，与三倍体cru鱼脑细胞和二倍体金鱼脑细胞相吻合。聚合酶链反应扩增和序列分析表明与已知cru鱼线粒体DNA序列的100％和94％匹配。接种了由塞浦路斯疱疹病毒2（CyHV-2）暴发的病态或死亡的金鱼组织匀浆接种后，在两种细胞系中连续观察了超过10代的细胞病变效应。这些新建立的细胞系可能是鱼类病毒性疾病的诊断工具。接种了由塞浦路斯疱疹病毒2（CyHV-2）暴发的病态或死亡的金鱼组织匀浆接种后，在两种细胞系中连续观察了超过10代的细胞病变效应。这些新建立的细胞系可能是鱼类病毒性疾病的诊断工具。接种了由塞浦路斯疱疹病毒2（CyHV-2）暴发的病态或死亡的金鱼组织匀浆接种后，在两种细胞系中连续观察了超过10代的细胞病变效应。这些新建立的细胞系可能是鱼类病毒性疾病的诊断工具。