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RNA Stability in Human Liver: Comparison of Different Processing Times, Temperatures and Methods

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Abstract

The accuracy of information garnered by real-time quantitative polymerase chain reaction (RT-qPCR), an important technology for elucidating molecular mechanisms of disease, is dependent on tissue quality. Thus, this study aimed to determine the effects of intra-operative manipulation, extended processing times, different temperatures or storage in RNAlater on RNA quality in liver samples for tissue banking. Liver samples, flash-frozen or in RNAlater, were collected over a time course (during surgery before blood arrest up to 1 day after surgery) with samples kept either at room temperature (RT) or on ice. This study showed that at the longest time-point at RT, the RNA quality decreased significantly by 20%. However, relative gene expressions of FOS, GUSB, MYC, HIF1α and GFER were in general not significantly different when the time-points were compared. In conclusion, samples should be kept on ice during processing, and either RNAlater or snap-freezing should be utilised for storage. Further, intra-operative manipulation and extended postoperative processing time generally does not change relative gene expression levels for the 5 genes studied, making such sampling suitable for RT-qPCR analysis. Thus, if relative gene expression of a gene of interest is stable, these guidelines will lead to increased accrual of samples to the tissue bank.

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Acknowledgments

This work was made possible by the HTCR foundation, which makes human tissues available for research. We thank the technical assistants from the Grosshadern Hospital Tissue Bank for collecting the liver samples. Our thanks also go to Dr. Bauersachs from the Gene Center in Ludwig Maximilians University for kindly allowing us to use his Agilent Bioanalyzer. This work was carried out with financial support from the Bundesministerium für Bildung und Forschung (BMBF; grant name: M4 Munich Biotech Excellence Cluster Biobank Alliance, grant number 01EX1020B).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Grosshadern Hospital Tissue Bank, Department of Surgery, Grosshadern Hospital, Ludwig Maximilians University, Munich, Germany

    Serene M. L. Lee, Celine Schelcher, Sevdije Gashi, Stefanie Schreiber & Wolfgang E. Thasler

  2. BioM BioTech Cluster Development GmbH, Martinsried, Germany

    Reinhard M. K. Thasler

  3. Department of Surgery, Grosshadern Hospital, Ludwig Maximilians University, Munich, Germany

    Karl-Walter Jauch & Wolfgang E. Thasler

Authors
  1. Serene M. L. Lee
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  2. Celine Schelcher
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  3. Sevdije Gashi
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  4. Stefanie Schreiber
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  5. Reinhard M. K. Thasler
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  6. Karl-Walter Jauch
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  7. Wolfgang E. Thasler
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Correspondence to Wolfgang E. Thasler.

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Serene M. L. Lee and Celine Schelcher are equal authors.

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Lee, S.M.L., Schelcher, C., Gashi, S. et al. RNA Stability in Human Liver: Comparison of Different Processing Times, Temperatures and Methods. Mol Biotechnol 53, 1–8 (2013). https://doi.org/10.1007/s12033-011-9493-4

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  • DOI: https://doi.org/10.1007/s12033-011-9493-4

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