Lässer Cecilia
Krefting Research Centre, Department of Internal Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Methods Mol Biol. 2013;1024:109-28. doi: 10.1007/978-1-62703-453-1_9.
Exosomes are 40-100 nm sized vesicles released from cells when multivesicular bodies fuse with the plasma membrane. These vesicles take part in cell-to-cell communication by binding and signalling through membrane receptors on cells or by transferring proteins, RNA, and lipids into the cells. Exosomal RNA in body fluids, such as plasma and urine, has been associated with malignancies, making the exosomal RNA a potential biomarker for early detection of these diseases. This has increased the interest in the field of extracellular RNA and in particular, the interest in exosomal RNA.In this chapter, a well-established exosome isolation method is described, as well as how to characterize the isolated vesicles by electron microscopy. Furthermore, two types of RNA isolation methods are described with a focus on isolating RNA from body fluids, which can be more viscous than cell culture media.
外泌体是当多泡体与质膜融合时从细胞中释放出来的大小为40 - 100纳米的囊泡。这些囊泡通过与细胞上的膜受体结合并发出信号,或通过将蛋白质、RNA和脂质转移到细胞中来参与细胞间通讯。体液(如血浆和尿液)中的外泌体RNA已被发现与恶性肿瘤有关,这使得外泌体RNA成为这些疾病早期检测的潜在生物标志物。这增加了人们对细胞外RNA领域的兴趣,尤其是对外泌体RNA的兴趣。在本章中,将描述一种成熟的外泌体分离方法,以及如何通过电子显微镜对分离出的囊泡进行表征。此外,还将描述两种RNA分离方法,重点是从比细胞培养基更粘稠的体液中分离RNA。