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Lee YXF, Johansson H, Wood MJA, et al. Considerations and Implications in the Purification of Extracellular Vesicles - A Cautionary Tale. Front Neurosci. 2019;13:1067doi: 10.3389/fnins.2019.01067.
Lee, Y. X. F., Johansson, H., Wood, M. J. A., & El Andaloussi, S. (2019). Considerations and Implications in the Purification of Extracellular Vesicles - A Cautionary Tale. Frontiers in neuroscience, 131067. https://doi.org/10.3389/fnins.2019.01067
Lee, Yi Xin Fiona, et al. "Considerations and Implications in the Purification of Extracellular Vesicles - A Cautionary Tale." Frontiers in neuroscience vol. 13 (2019): 1067. doi: https://doi.org/10.3389/fnins.2019.01067
Lee YXF, Johansson H, Wood MJA, El Andaloussi S. Considerations and Implications in the Purification of Extracellular Vesicles - A Cautionary Tale. Front Neurosci. 2019 Oct 18;13:1067. doi: 10.3389/fnins.2019.01067. eCollection 2019. PMID: 31680809; PMCID: PMC6813730.
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Front Neurosci. 2019 Oct 18;13:1067. doi: 10.3389/fnins.2019.01067. eCollection 2019.

Considerations and Implications in the Purification of Extracellular Vesicles - A Cautionary Tale.

Frontiers in neuroscience

Yi Xin Fiona Lee, Henrik Johansson, Matthew J A Wood, Samir El Andaloussi

Affiliations

  1. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom.
  2. Genome Institute of Singapore, Agency for Science, Technology and Research, Singapore, Singapore.
  3. Cancer Proteomics Mass Spectrometry, Science for Life Laboratory, Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
  4. Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.

PMID: 31680809 PMCID: PMC6813730 DOI: 10.3389/fnins.2019.01067

Abstract

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.

Copyright © 2019 Lee, Johansson, Wood and El Andaloussi.

Keywords: exosomes; extracellular vesicles; isolation; proteomics; purification; stem cells

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