Abuja Peter M, Ehrhart Friederike, Schoen Uwe, Schmidt Tomm, Stracke Frank, Dallmann Guido, Friedrich Torben, Zimmermann Heiko, Zatloukal Kurt
†Institute of Pathology, Medical University of Graz, Austria Auenbruggerplatz 25, A-8036 Graz, Austria.
‡Fraunhofer Institute of Biomedical Technology, Ensheimer Str. 48, 66386 St. Ingbert, Germany.
J Proteome Res. 2015 Jul 2;14(7):2758-68. doi: 10.1021/acs.jproteome.5b00025. Epub 2015 Jun 11.
Tissue metabolomics requires high sample quality that crucially depends on the biobanking storage protocol. Hence, we systematically analyzed the influence of realistic storage scenarios on the liver metabolome with different storage temperatures and repeated transfer of samples between storage and retrieval environments, simulating the repeated temperature changes affecting unrelated samples stored in the same container as the sample that is to be retrieved. By cycling between storage (-80 °C freezer, liquid nitrogen, cold nitrogen gas) and retrieval (room temperature, -80 °C), assuming three cycles per day and sample, we simulated biobank storage between 3 months and 10 years. Liver tissue metabolome was analyzed by liquid chromatography/mass spectrometry. Most metabolite concentrations changed <5% for the first "year" of time-compressed biobanking simulation, predominantly due to hydrolysis of peptides and lipids. Interestingly, storage temperature affected metabolite concentrations only little, while there was a linear dependence on the number of temperature change cycles. Elevated sample temperature during (prolonged) retrieval time led to a distinctly different signature of metabolite changes that were induced by cycling. Our findings allow giving recommendations for optimized storage protocols and provide signatures that allow detection of deviations from protocol.
组织代谢组学需要高质量的样本，而这在很大程度上取决于生物样本库的存储方案。因此，我们系统地分析了实际存储场景对肝脏代谢组的影响，研究了不同存储温度以及样本在存储和取回环境之间反复转移的情况，模拟了与待取回样本存于同一容器中的无关样本所受的反复温度变化影响。通过在存储（-80°C冰箱、液氮、冷氮气）和取回（室温、-80°C）之间循环，假设每天每个样本循环三次，我们模拟了生物样本库3个月至10年的存储情况。采用液相色谱/质谱法分析肝脏组织代谢组。在时间压缩的生物样本库模拟的第一个“年”中，大多数代谢物浓度变化<5%，主要原因是肽和脂质的水解。有趣的是，存储温度对代谢物浓度影响很小，而代谢物浓度与温度变化循环次数呈线性相关。（长时间）取回期间样本温度升高导致循环诱导的代谢物变化特征明显不同。我们的研究结果可为优化存储方案提供建议，并提供可检测方案偏差的特征。
