Andreasson Adam, Kiss Nimrod B, Juhlin C Christofer, Höög Anders
Biopreserv Biobank. 2013 Dec;11(6):366-70. doi: 10.1089/bio.2013.0038.
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°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.
To investigate to what extent long-term storage of tissues in -80°C affects the RNA quality and overall histomorphology. The tissue storage period represents nearly three decades (1986-2013).
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.
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.
Long-term storage in -80°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.
如今,关于如何最佳地保存人体组织以供长期储存尚无共识。液氮中的超低温储存常被推崇为长期保存的最佳方法,但在 -80 摄氏度(-80°C)冰箱中储存虽有时存在争议,却是一种可能的替代方法。在分子生物学背景下,RNA 是生物样本中最易降解的成分,其质量能够可靠地进行测定。
研究在 -80°C 下长期储存组织对 RNA 质量和整体组织形态学的影响程度。组织储存期将近三十年(1986 - 2013 年)。
使用 mirVana 试剂盒(Invitrogen)从 153 个不同储存期的组织样本中提取 RNA。使用安捷伦生物分析仪评估 RNA 完整性以获得 RNA 完整性数值(RIN)。此外,由两名病理学家通过光学显微镜进行组织代表性检测,以评估组织组成和形态。
对所有样本测量了 RIN 值,显示出的变异性与组织储存时间无关。在显微镜下,无论储存时间如何,所有样本均呈现出可接受的组织形态。
在 -80°C 下长期储存不会对从储存组织中提取的 RNA 质量产生不利影响,并且组织形态保持在良好标准。