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Size-Selective Harvesting of Extracellular Vesicles for Strategic Analyses Towards Tumor Diagnoses

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Abstract

Extracellular vesicles (EV), typified by exosomes or microvesicles, are expected to be effective diagnostic markers for cancers. The sizes of the vesicles range from 20 to 1000 nm, but the size-dependent variations of the contents of EVs are still poorly understood. We succeeded in the size-selective harvesting of the vesicles by utilizing the molecular weight-dependent characteristics of a variety of polyethylene glycols (PEG) as precipitating reagents and analyzed the antigens displayed on the surfaces of the vesicles and the miRNAs included in the vesicles from each size group. As a result, the relatively larger (<100 nm) particles precipitated by PEG5k clearly exhibited the greatest amount of epithelial cell adhesion molecule (EpCAM), from both breast cancer (MCF-7) and colon cancer (HCT116) cells, and a larger quantity of microRNA (miRNA) specific to breast cancer cells (miRNA155 for MCF-7) seemed to be contained in the PEG-precipitated particles. The results demonstrated that the quantities of both the tumor-specific miRNA and protein were similarly distributed among the several classes of the size-sorted EVs and that the size-selective harvesting of EVs may be informative for strategic analyses towards the diagnoses of cancers.

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Acknowledgments

The authors are grateful to Drs. T. Sakamoto and T. Takakura at Hitachi, Ltd. for helpful discussions and Ms. I. Makino and Ms. M. Shoji at Hitachi, Ltd. for experimental assistance.

Authors’ Contributions

CM and HN optimized and performed the size-selective separation of particles and performed the light scattering measurement experiments. CM performed all ELISA experiments and quantitative real-time PCR experiments. TY provided the starting materials and protocols for ELISA experiments and helped to write the manuscript. CM and HN prepared the figures and wrote the manuscript. All authors commented on the manuscript and approved the final version.

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Authors and Affiliations

  1. Center for Technology Innovation—Healthcare, Hitachi Ltd., Research & Development Group, 1-280 Higashi-koigakubo, Kokubunji, Tokyo, 185-8601, Japan

    Chihiro Manri, Takahide Yokoi & Hirokazu Nishida

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  1. Chihiro Manri
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  2. Takahide Yokoi
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  3. Hirokazu Nishida
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Correspondence to Hirokazu Nishida.

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Manri, C., Yokoi, T. & Nishida, H. Size-Selective Harvesting of Extracellular Vesicles for Strategic Analyses Towards Tumor Diagnoses. Appl Biochem Biotechnol 182, 609–623 (2017). https://doi.org/10.1007/s12010-016-2348-5

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  • DOI: https://doi.org/10.1007/s12010-016-2348-5

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