Abstract
Molecular investigations are crucial for further developments in precision medicine. RNA sequencing, alone or in combination with further omic-analyses, resulted in new therapeutic strategies. In this context, biobanks represent infrastructures to store tissue samples and body fluids in combination with clinical data to promote research for new predictive and prognostic biomarkers as well as therapeutic candidate molecules. Until today, the optimal storage conditions are a matter of debate especially with view to the storage temperature. In this unique approach we compared parallel samples from the same tumour, one half stored at − 80 °C and one half in the vapor phase of liquid nitrogen, with almost identical pre-analytical conditions. We demonstrated that RNA isolated from breast cancer samples revealed significantly higher RINe-values after 10 years of storage in the vapor phase of liquid nitrogen compared to storage at − 80 °C. In contrast, no significant difference was found regarding the DIN-values after DNA isolation. Morphological changes of the nucleus and cytoplasm, especially in the samples stored at − 80 °C, gave insights to degenerative effects, most possibly due to the storage protocol and its respective peculiarities. In addition, our results indicate that exact point-to point documentation beginning at the sample preparation is mandatory.
Highlights
Molecular investigations are crucial for further developments in precision medicine
This study aims to compare RIN equivalents (RINe, algorithm based calculation, Agilent TapeStation 4200)- and DIN-values from the same tumour tissue stored for 10 years under two different storage temperatures, namely under − 80 °C and in the VPLN corresponding to about − 186°C25
Correlating our R INe values to the four RNA “fit-for-purpose” quality groups stated by Kap et al our results can be split up as follows[16]: from the total of 16 specimens stored at − 80 °C, one sample was below the cut-off of < 5 and not reliable for downstream analysis. RINe-values of two samples ranged between ≥ 5 and < 6. RINe-values of eight samples ranged between ≥ 6 and < 8. RINe-values ≥ 8 were found in five samples
Summary
Molecular investigations are crucial for further developments in precision medicine. RNA sequencing, alone or in combination with further omic-analyses, resulted in new therapeutic strategies. In precision medicine and modern pathological diagnostics, molecular methods like gene expression analyses become more and more important to determine the origin of human diseases and to clear-up prognostic as well as predictive factors and to find appropriate new therapeutic strategies[1,2] In this context, the integrity of different omic-levels, namely the genome, epigenome, transcriptome, proteome and metabolome are of special interest to understand specific cellular mechanisms, which could be a target for clinical s trategies[3]. This study aims to compare RIN equivalents (RINe, algorithm based calculation, Agilent TapeStation 4200)- and DIN-values from the same tumour tissue stored for 10 years under two different storage temperatures, namely under − 80 °C and in the VPLN corresponding to about − 186°C25
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