Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)
Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)
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DOI：
10.1007/s10695-012-9733-y
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发表时间：
2013-06
影响因子：
2.9
通讯作者：
S. Higaki;Yoshie Koyama;E. Shirai;T. Yokota;Y. Fujioka;Noriyoshi Sakai;T. Takada
中科院分区：
文献类型：
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作者：
S. Higaki;Yoshie Koyama;E. Shirai;T. Yokota;Y. Fujioka;Noriyoshi Sakai;T. Takada
我们成功地建立了来自当地鱼类HonmorokoGnathopogon caerulescens的细胞系，这种鱼类栖息在世界第三古老的湖泊琵琶湖。从卵巢和精巢中分别建立了RMT 1和RMO 1细胞系。caerulescens，分别。这些细胞系最初在补充有胎牛血清（FBS）、鱼胚提取物、表皮生长因子和碱性成纤维细胞生长因子的Leibovitz L-15培养基中培养。需要进一步添加毛喉素和β-巯基乙醇以建立并维持这些细胞系超过60代。RMT 1和RMO 1细胞分别呈成纤维细胞样和上皮样形态。从免疫细胞化学染色和基因表达谱来看，RMT 1细胞具有睾丸支持细胞的特征，而RMO 1细胞具有卵巢卵泡膜细胞的特征。RMT 1和RMO 1细胞在28 °C下在补充有10%FBS的培养基中均能良好增殖，它们的最小群体倍增时间分别为24.4和28.8 h。在第45代时，大多数RMT 1和RMO 1细胞具有超倍体染色体组（分别为67.3%和96.1%）。未观察到具有正常二倍体染色体组的细胞。用增强型绿色荧光蛋白（EGFP）表达载体转染RMT 1细胞，人延伸因子1 α启动子可有效表达EGFP。此外，还建立了表达EGFP的细胞系，这表明该细胞系可用作体外监测系统（生物传感器），用于评价可能影响性腺功能的内分泌干扰物。
We succeeded to establish cell lines from endemic fish species HonmorokoGnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary ofG. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function.