Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples.
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 stor...
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Public Library of Science (PLoS)
2020-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0227648&type=printable |
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| author | Pamela R Matias-Garcia Rory Wilson Veronika Mussack Eva Reischl Melanie Waldenberger Christian Gieger Gabriele Anton Annette Peters Andrea Kuehn-Steven |
| author_facet | Pamela R Matias-Garcia Rory Wilson Veronika Mussack Eva Reischl Melanie Waldenberger Christian Gieger Gabriele Anton Annette Peters Andrea Kuehn-Steven |
| author_sort | Pamela R Matias-Garcia |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-c48dcb0b68b4422aa6d5e84b216bf058 |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj-art-c48dcb0b68b4422aa6d5e84b216bf0582025-08-20T02:54:53ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-01151e022764810.1371/journal.pone.0227648Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples.Pamela R Matias-GarciaRory WilsonVeronika MussackEva ReischlMelanie WaldenbergerChristian GiegerGabriele AntonAnnette PetersAndrea Kuehn-StevenSample 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0227648&type=printable |
| spellingShingle | Pamela R Matias-Garcia Rory Wilson Veronika Mussack Eva Reischl Melanie Waldenberger Christian Gieger Gabriele Anton Annette Peters Andrea Kuehn-Steven Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples. PLoS ONE |
| title | Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples. |
| title_full | Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples. |
| title_fullStr | Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples. |
| title_full_unstemmed | Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples. |
| title_short | Impact of long-term storage and freeze-thawing on eight circulating microRNAs in plasma samples. |
| title_sort | impact of long term storage and freeze thawing on eight circulating micrornas in plasma samples |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0227648&type=printable |
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