Martin Nicole M, Cooke Katherine M, Radford Caitlin C, Perley Lauren E, Silasi Michelle, Flannery Clare A
Department of Obstetrics and Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT, USA.
Am J Reprod Immunol. 2017 Apr;77(4). doi: 10.1111/aji.12637. Epub 2017 Jan 31.
Preservation of biospecimen quality is critical to accurately and reliably assessing genes and proteins. We evaluated the effect of preparation method and storage duration on RNA quality in placenta and decidua.
Aliquots of nine placentas and decidua were placed in RNAlater (RL) or flash frozen (FF) within 30 minutes of delivery. RNA was extracted immediately (baseline) and from matched samples stored at -80°C for 1 and 8-10 months. RNA Integrity Number (RIN) and housekeeping gene expression were quantified.
At both time points, RL placenta had RIN and housekeeping gene Ct values similar to baseline. However, FF placenta had significantly lower RIN and higher Ct values at 1 and 8-10 months. In RL and FF decidua, RIN was unchanged from baseline.
We found RNAlater more effectively and consistently preserved placenta, compared to flash freezing. However, for decidua, which is less dense than placenta, both modes yielded comparable RNA integrity over time.
生物样本质量的保存对于准确可靠地评估基因和蛋白质至关重要。我们评估了制备方法和储存时间对胎盘和蜕膜中RNA质量的影响。
在分娩后30分钟内，将9份胎盘和蜕膜的等分试样置于RNA Later（RL）中或速冻（FF）。立即提取RNA（基线），并从储存在-80°C下1个月和8 - 10个月的匹配样本中提取RNA。对RNA完整性数值（RIN）和管家基因表达进行定量分析。
在两个时间点，RL胎盘的RIN和管家基因Ct值与基线相似。然而，FF胎盘在1个月和8 - 10个月时RIN显著降低，Ct值升高。在RL和FF蜕膜中，RIN与基线无变化。
我们发现与速冻相比，RNA Later能更有效且一致地保存胎盘。然而，对于密度低于胎盘的蜕膜，随着时间推移，两种方式产生的RNA完整性相当。
