ISOENZYME ELECTROPHORESIS AND KARYOTYPE TECHNIQUES AS AN ALTERNATIVE FOR CELL LINE IDENTITY CONFIRMATION

J. Machado, A. Miranda, C. R. Gonçalves, T. I. Ikeda, Á. S. Cruz
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引用次数: 2

Abstract

The use of cell culture in biomedical science has been constantly enhanced and the use of certified and microrganisms-free cell lines is indispensable. Due to the value and the difficulty of acquiring a commercially available kit (AuthentikitTM system) for our routine in addition with the purpose of monitoring and ensuring quality of provided cell lines, and also contributing with cell characterization, the Nucleo de Cultura de Celulas do Instituto Adolfo Lutz (NCC– AL) padronized two techniques to identify the animal species from which the cells were originated: isoenzyme electrophoresis and karyotype. Cell extracts from 14 different cell lines were run under horizontal electrophoresis in agarose gel and lactate dehydrogenase isoenzyme profile was revealed using tetrazolium salt. The number of bands and the migration distances between bands of cell lines were similar to the literature, but small adjustments were needed to assist in the visualization of the bands that were not visible. Cell flasks for karyotype were incubated with colcemid solution and modal values of each cell line were calculated. Comparing all the results obtained with those reported in the literature, we conclude that both methods are promising and will allow better control of the services provided by the laboratory, not to mention guaranteeing authenticated cell lines for research and diagnostics. DOI:  http://dx.doi.org/10.17525/vrr.v19i1.90
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同工酶电泳和核型技术作为细胞系身份确认的替代方法
细胞培养在生物医学科学中的应用不断加强,使用经过认证和无微生物的细胞系是必不可少的。由于获得商用试剂盒(AuthentikitTM系统)的价值和难度,此外,为了监测和确保所提供细胞系的质量,并有助于细胞表征,Adolfo Lutz研究所(NCC - AL)的细胞核培养(Nucleo de Cultura de Celulas do Instituto Adolfo Lutz)批准了两种技术来鉴定细胞起源的动物物种:同工酶电泳和核型。用琼脂糖凝胶对14个不同细胞系的细胞提取物进行水平电泳,并用四氮唑盐测定乳酸脱氢酶同工酶谱。细胞系的条带数量和条带之间的迁移距离与文献相似,但需要进行小的调整,以帮助可视化不可见的条带。用秋碱溶液孵育核型细胞瓶,计算各细胞系的模态值。将获得的所有结果与文献报道的结果进行比较,我们得出结论,这两种方法都是有希望的,并且可以更好地控制实验室提供的服务,更不用说为研究和诊断保证经过认证的细胞系了。DOI: http://dx.doi.org/10.17525/vrr.v19i1.90
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