Long-Term Storage of Endocrine Tissues at-80°C Does Not Adversely Affect RNA Quality or Overall Histomorphology
Long-Term Storage of Endocrine Tissues at-80°C Does Not Adversely Affect RNA Quality or Overall Histomorphology
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DOI：
10.1089/bio.2013.0038
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发表时间：
2013-12-01
影响因子：
1.6
通讯作者：
Hoog, Anders
中科院分区：
文献类型：
--
作者：
Andreasson, Adam;Kiss, Nimrod B.;Hoog, Anders
背景：今天，关于如何最好地保存人体组织以供长期储存，还没有达成共识。液氮中的极低温储存通常被认为是延长时间的最佳方法，但储存在-80摄氏度（-80摄氏度）的冷冻机中，虽然有时有争议，但也是一种可能的替代方法。在分子生物学背景下，RNA是生物样品中最容易降解的组分，并且可以可靠地测量其质量。目的：研究组织在-80 ℃下长期储存对RNA质量和整体组织形态学的影响程度。组织储存期为近三十年（1986-2013）。方法：采用mirVana试剂盒（Invitrogen）从153份不同保存时间的组织样本中提取RNA。使用Agilent生物分析仪评估RNA完整性以获得RNA完整性编号（RIN）。此外，由两名病理学家使用光学显微镜进行组织代表性试验，以评估组织组成和形态。结果：在所有样本中测量RIN值，显示与组织储存时间无关的变异性。在显微镜下，所有样品均显示出可接受的组织形态，与储存时间无关。结论：在-80 ℃下长期储存不会对从储存的组织中提取的RNA的质量产生不利影响，并且组织形态保持良好的标准。
Background: Today, no consensus exists regarding how human tissues are best preserved for long-term storage. Very low temperature storage in liquid nitrogen is often advocated as the superlative method for extended periods, but storage in -80 degrees Celsius (-80 degrees C) freezers, while sometimes debated, is a possible alternative. RNA is the most easily degradable component of a biological sample in a molecular biology context and the quality can reliably be measured. Aim: To investigate to what extent long-term storage of tissues in -80 degrees C affects the RNA quality and overall histomorphology. The tissue storage period represents nearly three decades (1986-2013). Methods: RNA extraction from 153 tissue samples with different storage periods was performed with the mirVana kit (Invitrogen). RNA integrity was assessed using an Agilent bioanalyzer to obtain RNA integrity numbers (RIN). Further, tissue representative testing using light microscopy was performed by two pathologists to assess tissue composition and morphology. Results: RIN values were measured in all samples, showing a variability that did not correlate with the storage time of the tissues. Microscopically, all samples displayed acceptable tissue morphology regardless of storage time. Conclusion: Long-term storage in -80 degrees C does not adversely affect the quality of the RNA extracted from the stored tissues, and the tissue morphology is maintained to a good standard.