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对安达卢西亚公共生物样本库实施的长期冷冻保存和核酸提取过程的质量评估。

Quality assessment on the long-term cryopreservation and nucleic acids extraction processes implemented in the andalusian public biobank.

作者信息

Ortega-Pinazo J, Díaz T, Martínez B, Jiménez A, Pinto-Medel M J, Ferro P

机构信息

Neuroscience UGC, Instituto de Investigación Biomédica de Málaga (IBIMA), Sanitary Distric of Málaga, University of Málaga, Málaga, Spain.

Andalusian Public Health System Biobank, Instituto de Investigación Biomédica de Málaga (IBIMA), Sanitary Distric of Málaga, University of Málaga, Málaga, Spain.

出版信息

Cell Tissue Bank. 2019 Jun;20(2):255-265. doi: 10.1007/s10561-019-09764-9. Epub 2019 Mar 22.

Abstract

Human samples are commonly collected and long-term stored in biobanks for current and future analyses. Even though techniques for freezing human blood are well established, the storage time can compromise the cell viability as well as the yield and quality of nucleic acids (RNA and DNA) extracted from them. In this study, a protocol to obtain peripheral blood mononuclear cells (PBMCs) from 70 subjects, which were stored at - 196 °C from EDTA tubes for a long-term, was assessed. In parallel; a protocol to obtain DNA from the same subjects, which were stored at - 80 °C from citrate tubes, was also studied. Samples stored from 2008 to 2012 were studied and the results obtained showed that there were no statistically significant differences in the RNA or DNA extracted in terms of purity, integrity and functionality The freezing protocol used by the Málaga Biobank shows that viable PBMCs and DNA could be kept for a period of, at least, 10 years, with a high quality and performance. Furthermore, RNA extracted from these PBMCs presents also a good quality and performance. Therefore, the samples frozen according to the conditions of the protocols assessed in this study could be optimal for biomedical research.

摘要

人类样本通常被采集并长期储存在生物样本库中,以供当前和未来的分析使用。尽管冷冻人类血液的技术已经很成熟,但储存时间可能会影响细胞活力以及从中提取的核酸(RNA和DNA)的产量和质量。在本研究中,评估了一种从70名受试者中获取外周血单核细胞(PBMC)的方案,这些样本长期保存在-196°C的EDTA管中。同时,还研究了一种从同一受试者中获取DNA的方案,这些样本保存在-80°C的柠檬酸盐管中。对2008年至2012年储存的样本进行了研究,结果表明,在提取的RNA或DNA的纯度、完整性和功能性方面没有统计学上的显著差异。马拉加生物样本库使用的冷冻方案表明,有活力的PBMC和DNA可以至少保存10年,且质量和性能良好。此外,从这些PBMC中提取的RNA也具有良好的质量和性能。因此,根据本研究评估的方案条件冷冻的样本可能是生物医学研究的最佳选择。

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