Tosar Juan Pablo, Cayota Alfonso, Eitan Erez, Halushka Marc K, Witwer Kenneth W
Functional Genomics Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay; Nuclear Research Center, Faculty of Science, Universidad de la República, Montevideo, Uruguay.
Functional Genomics Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay; Department of Medicine, Faculty of Medicine, Universidad de la República, Montevideo, Uruguay.
J Extracell Vesicles. 2017 Jan 12;6(1):1272832. doi: 10.1080/20013078.2016.1272832. eCollection 2017.
In a recently published study, Anna Krichevsky and colleagues raise the important question of whether results of in vitro extracellular RNA (exRNA) studies, including extracellular vesicle (EV) investigations, are confounded by the presence of RNA in cell culture medium components such as foetal bovine serum (FBS). The answer, according to their data, is a resounding "yes". Even after lengthy ultracentrifugation to remove bovine EVs from FBS, the majority of exRNA in FBS remained. Although technical factors may affect the degree of depletion, residual EVs and exRNA in FBS could influence the conclusions of in vitro studies: certainly, for secreted RNA, and possibly also for cell-associated RNA. In this commentary, we critically examine some of the literature in this field, including a recent study from some of the authors of this piece, in light of the Wei et al. study and explore how cell culture-derived RNAs may affect what we think we know about EV RNAs. These findings hold particular consequence as the field moves towards a deeper understanding of EV-RNA associations and potential functions.
在最近发表的一项研究中，安娜·克里切夫斯基及其同事提出了一个重要问题：包括细胞外囊泡（EV）研究在内的体外细胞外RNA（exRNA）研究结果，是否会受到细胞培养基成分（如胎牛血清，FBS）中RNA的干扰。根据他们的数据，答案是响亮的“是”。即使经过长时间超速离心以从FBS中去除牛源细胞外囊泡，FBS中的大多数exRNA仍然存在。尽管技术因素可能会影响去除程度，但FBS中残留的细胞外囊泡和exRNA可能会影响体外研究的结论：当然，对于分泌型RNA是这样，对于细胞相关RNA可能也是如此。在这篇评论中，我们根据魏等人的研究，批判性地审视了该领域的一些文献，包括本文部分作者最近的一项研究，并探讨了细胞培养衍生的RNA如何可能影响我们对细胞外囊泡RNA的认知。随着该领域朝着更深入理解细胞外囊泡 - RNA关联及潜在功能发展，这些发现具有特别重要的意义。
