Wang Min, Ji Xiaoli, Wang Bingjie, Li Qian, Zhou Junmei
Department of Central Laboratory, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China.
Biopreserv Biobank. 2018 Dec;16(6):426-433. doi: 10.1089/bio.2018.0055. Epub 2018 Nov 28.
A major concern in biomedical research is the quality of biological samples. RNAlater is a stabilizer, which was originally developed for RNA preservation in fresh tissues and is important for collection and transportation. However, this reagent lacks a comprehensive and systematic evaluation of its preservative effect on different mammalian tissues under consistent experimental conditions. In this study, we collected liver, kidney, testis, brain, and colon tissues from mice and divided the samples into the following respective groups: fresh, RNAlater preserved, and liquid nitrogen snap frozen. Biomolecules (RNA, DNA, and protein) were extracted from each tissue in each group, and samples were formalin fixed and paraffin embedded for quality assessment. Our results revealed that high-quality (yield, purity, and integrity) nucleic acids could be extracted from all samples. Gene expression determined by quantitative real-time polymerase chain reaction exhibited no major difference among the three groups. Notably, we observed significant protein degradation in brain tissue preserved by RNAlater compared with fresh and snap-frozen tissue. Protein expression of the other four tissues was similar among the three groups. Hematoxylin and eosin staining of all tissue types indicated no apparent difference among the three groups. We concluded that high-quality nucleic acids can be obtained and tissue morphology conserved when tissues are preserved with RNAlater. However, there are tissue-specific differences in protein preservation when using RNAlater, which should be evaluated before extensive storage.
生物医学研究中的一个主要关注点是生物样本的质量。RNA later是一种稳定剂，最初是为在新鲜组织中保存RNA而开发的，对样本的采集和运输很重要。然而，该试剂在一致的实验条件下，缺乏对其在不同哺乳动物组织中的保存效果进行全面系统的评估。在本研究中，我们从小鼠身上采集了肝脏、肾脏、睾丸、大脑和结肠组织，并将样本分为以下几组：新鲜组、RNA later保存组和液氮速冻组。从每组的每个组织中提取生物分子（RNA、DNA和蛋白质），并将样本用福尔马林固定、石蜡包埋以进行质量评估。我们的结果表明，所有样本均可提取出高质量（产量、纯度和完整性）的核酸。通过定量实时聚合酶链反应测定的基因表达在三组之间没有显著差异。值得注意的是，与新鲜组和速冻组相比，我们观察到用RNA later保存的脑组织中蛋白质有明显降解。其他四种组织的蛋白质表达在三组之间相似。所有组织类型的苏木精和伊红染色显示三组之间没有明显差异。我们得出结论，当用RNA later保存组织时，可以获得高质量的核酸并保持组织形态。然而，使用RNA later保存蛋白质时存在组织特异性差异，在大量保存之前应进行评估。
