10.3389/fnins.2019.01067.s001
Yi Xin Fiona Lee
Yi Xin Fiona
Lee
Henrik Johansson
Henrik
Johansson
Matthew J. A. Wood
Matthew J. A.
Wood
Samir El Andaloussi
Samir El
Andaloussi
Image_1_Considerations and Implications in the Purification of Extracellular Vesicles – A Cautionary Tale.JPEG
Frontiers
2019
extracellular vesicles
stem cells
proteomics
exosomes
isolation
purification
2019-10-18 12:39:01
Figure
https://frontiersin.figshare.com/articles/figure/Image_1_Considerations_and_Implications_in_the_Purification_of_Extracellular_Vesicles_A_Cautionary_Tale_JPEG/10002536
<p>Extracellular vesicles (EVs) are nano-sized particles constitutively released from cells into all biological fluids. Interestingly, these vesicles contain genetic cargoes including proteins, RNA and bioactive lipids that can be functionally delivered and affect recipient cells. As a result, there is growing interest in studying EVs in pathological conditions, including central nervous system (CNS)-related diseases, as EVs may be used for diagnostic purposes or as therapeutic agents. However, one major bottleneck is the need for better EV purification strategies when considering complex biological sources such as serum/protein-rich media or plasma. In this study, we have performed a systematic comparison study between the current gold-standard method: ultracentrifugation, to an alternative: size-exclusion chromatography (LC), using induced pluripotent stem cell (iPSC) derived complex media as a model system. We demonstrate that LC allows for derivation of purer EVs from iPSCs, which was previously impossible with the original UC method. Importantly, our study further highlights the various drawbacks when using the conventional UC approach that lead to misinterpretation of EV data. Lastly, we describe novel data on our iPSC-EVs; how they could relate to stem cell biology and discuss their potential use as EV therapeutics for CNS diseases.</p>