Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples

Autoři: Pamela R. Matias-Garcia aff001;  Rory Wilson aff001;  Veronika Mussack aff004;  Eva Reischl aff001;  Melanie Waldenberger aff001;  Christian Gieger aff001;  Gabriele Anton aff001;  Annette Peters aff002;  Andrea Kuehn-Steven aff001
Působiště autorů: Research Unit of Molecular Epidemiology, Helmholtz Zentrum Muenchen, German Center for Environmental Health, Neuherberg, Germany aff001;  Institute of Epidemiology, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health, Neuherberg, Germany aff002;  TUM School of Medicine, Technical University of Munich, Munich, Germany aff003;  Department of Animal Physiology and Immunology, TUM School of Life Sciences Weihenstephan, Technical University of Munich (TUM), Freising, Germany aff004;  German Center for Cardiovascular Research (DZHK), partner site Munich Heart Alliance, Munich, Germany aff005;  German Center for Infection Research (DZIF), partner site Munich, Munich, Germany aff006
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article


Sample collection, processing, storage and isolation methods constitute pre-analytic factors that can influence the quality of samples used in research and clinical practice. With regard to biobanking practices, a critical point in the sample’s life chain is storage, particularly long-term storage. Since most studies examine the influence of different temperatures (4°C, room temperature) or delays in sample processing on sample quality, there is only little information on the effects of long-term storage at ultra-low (vapor phase of liquid nitrogen) temperatures on biomarker levels. Among these biomarkers, circulating miRNAs hold great potential for diagnosis or prognosis for a variety of diseases, like cancer, infections and chronic diseases, and are thus of high interest in several scientific questions. We therefore investigated the influence of long-term storage on levels of eight circulating miRNAs (miR-103a-3p, miR-191-5p, miR-124-3p, miR-30c-5p, miR-451a, miR-23a-3p, miR-93-5p, miR-24-3p, and miR-33b-5p) from 10 participants from the population-based cohort study KORA. Sample collection took place during the baseline survey S4 and the follow-up surveys F4 and FF4, over a time period spanning from 1999 to 2014. The influence of freeze-thaw (f/t) cycles on miRNA stability was also investigated using samples from volunteers (n = 6). Obtained plasma samples were profiled using Exiqon’s miRCURYTM real-time PCR profiling system, and repeated measures ANOVA was used to check for storage or f/t effects. Our results show that detected levels of most of the studied miRNAs showed no statistically significant changes due to storage at ultra-low temperatures for up to 17 years; miR-451a levels were altered due to contamination during sampling. Freeze-thawing of one to four cycles showed an effect only on miR-30c-5p. Our results highlight the robustness of this set of circulating miRNAs for decades of storage at ultra-low temperatures and several freeze-thaw cycles, which makes our findings increasingly relevant for research conducted with biobanked samples.

Klíčová slova:

Biomarkers – Blood plasma – Data visualization – MicroRNAs – Oligonucleotides – Platelets – principal component analysis – Specimen storage


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