Human blood RNA stabilization in samples collected and transported for a large biobank

Duale, Nur; Brunborg, Gunnar; Rønningen, Kjersti S.; Briese, Thomas; Aarem, Jeanette; Aas, Kaja K; Magnus, Per; Stoltenberg, Camilla; Süsser, Ezra; Lipkin, W. Ian

doi:10.1186/1756-0500-5-510

Cited by 47 publications

(71 citation statements)

References 18 publications

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“…Consistent with our recent report on effects of sample collection method and short term storage of RNA samples [4], the yields in cord blood samples were significantly higher than in the adult blood samples (data not shown). This difference is likely to reflect the higher cell content of the cord blood [11].…”

Section: Resultssupporting

confidence: 92%

Long-term storage of blood RNA collected in RNA stabilizing Tempus tubes in a large biobank – evaluation of RNA quality and stability

et al. 2014

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BackgroundEstablishing methods for secure long term storage of RNA is critical to realizing the promise of biobanks in biomedical research. Here, we describe the results of yearly analyses of the same set of umbilical cord and adult whole blood RNA collected in Tempus Blood RNA tubes and stored at -80°C, over a period of up to six years. We systematically investigated the effects of long-term storage of samples (75 Tempus tubes form three adult donors and 30 Tempus tubes from three cord blood donors) on the RNA quality and transcript stability of six selected genes (CDKN1A, FOS, IL1B, IL8, MYC and TP53). This is the first systematic study of both cord and adult blood samples stored for many years.FindingsThe RNA purity and integrity, expressed as RIN-values, were stable up to six years of storage, and there were no storage-related deleterious effects on RNA purity. There were limited intra- and inter-individual variations in RNA yields; however, no consistent trend of decreasing RNA yield was observed with the duration of storage. Some long-term storage effects were found on the relative transcript levels of the six genes when compared to the year 0 samples. However, these changes were within ± 2–fold for both types of blood samples, except for two genes. Our results show that storage of these samples for up to six years did not have significant effects on the RNA quality and transcript stability of the six genes.ConclusionsBlood RNA is stable in Tempus tubes stored at -80°C over a period of six years. Intact and good-quality RNA suitable for transcript profiling analyses in epidemiological studies was obtained from blood samples stored in Tempus tubes. This suggests that blood samples collected in large biobanks–such as the Mother and Child (MoBa) Cohort at Norwegian Institute of Public Health (NIPH) and frozen in suitable collection tubes for total RNA stabilization, can be used for quantitative studies after at least six years of storage.Electronic supplementary materialThe online version of this article (doi:10.1186/1756-0500-7-633) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 92%

Long-term storage of blood RNA collected in RNA stabilizing Tempus tubes in a large biobank – evaluation of RNA quality and stability

et al. 2014

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“…The results obtained by participants using the column-based methods were consistent with previous data showing that similar RNA quality, in terms of purity and RIN, was obtained independently from the extraction method (silica membrane vs. magnetic beads). 33,34 FFPE cells. DNA extracted from FFPE tissues is suitable for PCR, microarray, and sequencing analyses, although it is often difficult to amplify long fragments due to DNA fragmentation and cross-linking.…”

Section: Comparison Of Processing Methods: Nucleic Acid Extractionmentioning

confidence: 99%

A Biospecimen Proficiency Testing Program for Biobank Accreditation: Four Years of Experience

Gaignaux

Ashton

Coppola

et al. 2016

Biopreservation and Biobanking

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Biobanks produce and distribute biospecimens, ensuring their fitness for purpose and accurately qualifying them before distribution. In their efforts toward professionalization, biobanks can nowadays seek certification or accreditation. One of the requirements of these standards is regular participation in Proficiency Testing (PT) programs. An international PT program has been developed and provided to biobanks and other laboratories that perform specific tests to qualify different types of biospecimens. This PT program includes biospecimen testing schemes, as well as biospecimen processing interlaboratory exercises. This PT program supports the development of biobank quality assurance by providing the possibility to assess biobank laboratory performance and useful insights into biobank laboratory method performance characteristics and thus fulfill the demands from accreditation authorities.

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“…Total RNA was isolated from fresh frozen liver and testis tissues as previously described (Duale et al ., , ) using the GenElute kit (Sigma‐Aldrich, Inc., Oslo, Norway). RNA quantity and purity were determined using a NanoDrop 1000 Spectrophotometer (Fisher Scientific, Oslo, Norway).…”

Section: Methodsmentioning

confidence: 99%

Enhanced susceptibility of obese mice to glycidamide‐induced sperm chromatin damage without increased oxidative stress

et al. 2016

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Diet-induced obesity is known to impair male reproduction and may aggravate the male reproductive toxicity of the food contaminant acrylamide. Exposure of male mice to acrylamide induces paternally mediated pre- and post-implantation losses because of spermatozoal toxicity and these effects are potentiated in mice fed a high-fat diet. Glycidamide - an acrylamide metabolite - is the primary mediator of reproductive effects in males. The mechanisms causing the interaction between diet and acrylamide are not clear. However, diet-induced obesity is associated with oxidative stress in male reproductive tissues which might contribute to increased germ cell susceptibility. In this study, we investigated whether a moderate diet-induced obesity regimen could interfere with glycidamide-induced spermatozoal toxicity and increase oxidative stress. For this purpose, sperm chromatin integrity, oxidised DNA and protein levels, transcript levels of oxidative stress responsive genes and glycidamide-induced DNA and haemoglobin adducts were analysed in samples from male mice exposed to a high-fat diet for 6 weeks in combination with a single glycidamide exposure 7 days prior to sacrifice. We found that glycidamide-induced sperm DNA fragmentation was markedly higher in obese than in lean mice. However, the levels of oxidised DNA and/or protein in blood, liver and testicular tissue was lower in obese than in lean mice. Accompanying the reduced level of oxidised macromolecules, the transcript levels of several oxidative stress-related genes were altered in the liver and testis from obese mice suggesting induction of an antioxidant response in these animals. The haemoglobin-glycidamide adduct levels were higher in obese than in lean animals, whereas obesity did not seem to increase the level of glycidamide-induced DNA adducts. These findings show that a moderate diet-induced obesity regimen may potentiate glycidamide-induced sperm cells toxicity and suggest that the increase in glycidamide-induced sperm toxicity observed in obese mice does not depend on overt oxidative stress.

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Human blood RNA stabilization in samples collected and transported for a large biobank

Cited by 47 publications

References 18 publications

Long-term storage of blood RNA collected in RNA stabilizing Tempus tubes in a large biobank – evaluation of RNA quality and stability

Long-term storage of blood RNA collected in RNA stabilizing Tempus tubes in a large biobank – evaluation of RNA quality and stability

A Biospecimen Proficiency Testing Program for Biobank Accreditation: Four Years of Experience

Enhanced susceptibility of obese mice to glycidamide‐induced sperm chromatin damage without increased oxidative stress

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