Department of Marine Biology, College of Marine Life Sciences, Ocean University of China, 266003, Qingdao, China,
Cytotechnology. 2010 Jul;62(3):217-23. doi: 10.1007/s10616-010-9281-1. Epub 2010 May 26.
A turbot, Scophthalmus maximus, fin (TF) cell line was established and susceptibility to turbot reddish body iridovirus (TRBIV) was determined in this study. Primary culture of TF cells was initiated from fin tissue pieces partially digested with trypsin, collagenase II and hyaluronidase. Digested tissue pieces were cultured at 24 degrees C in Leibovitz-15 medium (pH 7.2), supplemented with 20% fetal bovine serum, carboxymethyl chitosan, N-acetylglucosamine hydrochloride, basic fibroblast growth factor and epidermal growth factor. The cultured TF cells, in fibroblast shape, proliferated to 100% confluency 50 days later. A TF cell line, with a population doubling time of 45.6 h at passage 80, has been established and subcultured to passage 133. Chromosome analyses indicated that the TF cells exhibited chromosomal aneuploidy with a modal chromosome number of 44 which displayed the normal diploid karyotype of S. maximus at least up to passage 80. TRBIV susceptibility testing demonstrated that cytopathic effect and propagated viral particles were observed in TF cells after TRBIV infection. In conclusion, a continuous TRBIV susceptible TF cell line has been established successfully, and the cell line may serve as a valuable tool for studies of cell-virus interactions and has applications for different kinds of cytotechnological studies as well.
本研究建立了大菱鲆鳍(TF)细胞系,并测定了其对大菱鲆虹彩病毒(TRBIV)的敏感性。TF 细胞的原代培养是从部分用胰蛋白酶、胶原酶 II 和透明质酸酶消化的鳍组织块开始的。消化后的组织块在 24°C 的 Leibovitz-15 培养基(pH 7.2)中培养,补充 20%胎牛血清、羧甲基壳聚糖、N-乙酰氨基葡萄糖盐酸盐、碱性成纤维细胞生长因子和表皮生长因子。培养的 TF 细胞呈成纤维细胞形态,50 天后增殖至 100%汇合。建立了一个 TF 细胞系,在第 80 代的倍增时间为 45.6 小时,已经传代至 133 代。染色体分析表明,TF 细胞表现出染色体非整倍性,模式染色体数为 44,至少在第 80 代显示出大菱鲆的正常二倍体核型。TRBIV 敏感性测试表明,在 TRBIV 感染后,TF 细胞中观察到细胞病变效应和增殖的病毒颗粒。总之,成功建立了一种连续的 TRBIV 敏感 TF 细胞系,该细胞系可作为研究细胞-病毒相互作用的有用工具,并且在各种细胞技术研究中也有应用。