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Cell lines authentication and mycoplasma detection as minimun quality control of cell lines in biobanking

机译:细胞系鉴定和支原体检测作为生物库中细胞系的最低质量控制

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摘要

Establishment of continuous cell lines from human normal and tumor tissues is an extended and useful methodology for molecular characterization of cancer pathophysiology and drug development in research laboratories. The exchange of these cell lines between different labs is a common practice that can compromise assays reliability due to contamination with microorganism such as mycoplasma or cells from different flasks that compromise experiment reproducibility and reliability. Great proportions of cell lines are contaminated with mycoplasma and/or are replaced by cells derived for a different origin during processing or distribution process. The scientific community has underestimated this problem and thousand of research experiment has been done with cell lines that are incorrectly identified and wrong scientific conclusions have been published. Regular contamination and authentication tests are necessary in order to avoid negative consequences of widespread misidentified and contaminated cell lines. Cell banks generate, store and distribute cell lines for research, being mandatory a consistent and continuous quality program. Methods implementation for guaranteeing both, the absence of mycoplasma and authentication in the supplied cell lines, has been performed in the Andalusian Health System Biobank. Specifically, precise results were obtained using real time PCR detection for mycoplasma and 10 STRs identification by capillary electrophoresis for cell line authentication. Advantages and disadvantages of these protocols are discussed.
机译:从人类正常组织和肿瘤组织建立连续细胞系是在研究实验室中用于癌症病理生理学和药物开发的分子表征的扩展且有用的方法。在不同实验室之间交换这些细胞系是一种常见做法,由于微生物(如支原体)或来自不同烧瓶的细胞受到污染,从而损害了实验的可重复性和可靠性,因此会损害测定的可靠性。在处理或分配过程中,很大比例的细胞系被支原体污染和/或被衍生自不同来源的细胞所替代。科学界低估了这个问题,对错误识别的细胞系进行了成千上万的研究实验,并发表了错误的科学结论。定期进行污染和鉴定测试是必要的,以避免广泛错误识别和污染的细胞系的负面影响。细胞库生成,存储和分配用于研究的细胞系,这是强制性的,连续且连续的质量计划。在安达卢西亚卫生系统生物库中已经执行了用于保证所提供的细胞系中不存在支原体和认证的方法。具体而言,使用实时PCR检测支原体和通过毛细管电泳鉴定10个STRs进行细胞系鉴定可获得精确的结果。讨论了这些协议的优缺点。

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