外泌体之家 | 细胞外膜泡领域核心平台—exosomes & microvesicles—小膜泡大作用
标题: exosomes消除步骤在去除胎牛血清中胞外囊泡功能和RNA的重要性 [打印本页] 作者: yy8651 时间: 2015-6-9 00:05 标题: exosomes消除步骤在去除胎牛血清中胞外囊泡功能和RNA的重要性 请原谅我这蹩脚的英语吧，大家还是自己看吧 胞外囊泡杂志上实用的文章 Importance of exosome depletion protocols to eliminate functional and RNA-containing extracellular vesicles from fetal bovine serum
Extracellular vesicles (EVs), including the nano-sized exosomes, have the capacity to transfer multiple
functional molecules between cells. In cell culture experiments, fetal bovine serum (FBS) is often used to
supplement cell culture medium as a nutrient, but it is important to know that the FBS also contain significant
quantities of EVs. The aim of the current study was to determine whether the FBS EVs can influence cultured
cell phenotype, and secondly to determine the efficiency of FBS-EVelimination protocols. Firstly, FBS that had
not been depleted of EVs induced a migratory phenotype in a lung cancer epithelial cell line (A549 cells), an
effect that could be mimicked by isolated FBS EVs alone. FBS-derived EVs also contained RNA, which was
protected from consecutive proteinase K and RNase A treatment. Comparison of common isolation protocols
suggested that an 18-hour centrifugation period eliminates approximately 95% of RNA-containing FBS EVs,
whereas a 1.5-hour protocol is insufficient. In conclusion, this study shows that FBS EVs substantially influence
cultured cell behaviour, but also that they can be virtually removed by an 18-hour ultracentrifugation protocol.
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