Béjar Julia, Porta Javier, Borrego J José, Alvarez M Carmen
Department of Genetics, Faculty of Sciences, University of Málaga, 29071, Málaga, Spain.
Mar Biotechnol (NY). 2005 Jul-Aug;7(4):389-95. doi: 10.1007/s10126-004-4083-0. Epub 2005 Jun 4.
Fish cell lines are increasingly important research tools. The SAF-1 cell line, fibroblast-like culture derived from the marine fish gilthead seabream (Sparus aurata), has proved useful in many applications, especially in viral research. For cell lines intended as in vitro models, characterization of their properties and authentication are essential for deeper understanding of their performance and thus more precise experimental design and applicability. In this study we characterized the SAF-1 cell line in terms of genetic stability through time and genetic labeling. Methods for determining stability include telomerase activity, karyotyping, mapping of ribosomal RNA regions, and DNA content. For genetic labeling 12 microsatellite loci were used. The results indicate that telomerase has been activated in the course of SAF-1 development, and the highest levels of telomerase activity correlate with an increase in cell proliferation, thus supporting a permanent cell line. This stability is in agreement with the normal situation presented by the cytogenetic traits and DNA content values, and the genotypic profile allows SAF-1 authentication at the single individual level. This study increases the value of SAF-1 as an in vitro system, which is now one of the few well-characterized cell lines from a marine fish.
鱼类细胞系正日益成为重要的研究工具。SAF-1细胞系是一种源自海鱼金头鲷(Sparus aurata)的成纤维样培养细胞系,已在许多应用中证明是有用的,特别是在病毒研究方面。对于用作体外模型的细胞系,表征其特性并进行鉴定对于更深入了解其性能,从而进行更精确的实验设计和应用至关重要。在本研究中,我们通过随时间的遗传稳定性和基因标记对SAF-1细胞系进行了表征。确定稳定性的方法包括端粒酶活性、核型分析、核糖体RNA区域图谱绘制和DNA含量测定。对于基因标记,使用了12个微卫星位点。结果表明,在SAF-1细胞系的发育过程中端粒酶被激活,端粒酶活性的最高水平与细胞增殖的增加相关,从而支持其作为永久细胞系。这种稳定性与细胞遗传学特征和DNA含量值所呈现的正常情况一致,并且基因型图谱允许在单个个体水平上对SAF-1进行鉴定。本研究提高了SAF-1作为体外系统的价值,它现在是少数几个特征明确的海鱼细胞系之一。