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Size and shape characterization of hydrated and desiccated exosomes

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Abstract

Exosomes are stable nanovesicles secreted by cells into the circulation. Their reported sizes differ substantially, which likely reflects the difference in the isolation techniques used, the cells that secreted them, and the methods used in their characterization. We analyzed the influence of the last factor on the measured sizes and shapes of hydrated and desiccated exosomes isolated from the serum of a pancreatic cancer patient and a healthy control. We found that hydrated exosomes are close-to-spherical nanoparticles with a hydrodynamic radius that is substantially larger than the geometric size. For desiccated exosomes, we found that the desiccated shape and sizing are influenced by the manner in which drying occurred. Isotropic desiccation in aerosol preserves the near-spherical shape of the exosomes, whereas drying on a surface likely distorts their shapes and influences the sizing results obtained by techniques that require surface fixation prior to analysis.

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Acknowledgments

The authors are indebted to Brian F. Woodfield of Brigham Young University (Department of Chemistry) for providing access to the NTA instrument. The authors acknowledge financial support from the National Science Foundation (award number IGERT-0903715) and the University of Utah (Department of Chemical Engineering Seed Grant and the Graduate Research Fellowship award).

Author contributions

M.S. and P.S.B. conceived the study, R.R. isolated exosomes, V.S.C. and R.R. performed NTA measurements, DLS measurements were performed by V.S.C. and M.S, D.M.B performed cryo-TEM imaging, SEM imaging was performed by Y.J. and V.S.C., electrospray DMA measurements were performed by Y.H.T., V.S.C., L.F.P., and M.S., V.S.C. and M.S. analyzed the experimental results; A.E.B. wrote the MATLAB code to analyze the imaged exosomes, M.S. performed statistical analysis, and V.S.C. and K.J.B. performed manual sizing of the imaged exosomes. The manuscript was written by V.S.C. and M.S., and was edited by all authors.

Conflict of interest

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Chemical Engineering, University of Utah, 50 Central Campus Drive, Salt Lake City, UT, 84112, USA

    Vasiliy S. Chernyshev, Yen Hsun Tseng, Yunlu Jia, Kyle J. Branch, Anthony E. Butterfield, Leonard F. Pease III & Mikhail Skliar

  2. Huntsman Cancer Institute, University of Utah, University Medical Center, 2000 Circle of Hope Drive, Salt Lake City, UT, 84112, USA

    Rakesh Rachamadugu & Philip S. Bernard

  3. Department of Biochemistry and Structural Biology, University of Utah, 257S 1400 E, Salt Lake City, UT, 84112, USA

    David M. Belnap

  4. Division of Gastroenterology, Hepatology, and Nutrition, Department of Internal Medicine, University of Utah, Salt Lake City, UT, 84112, USA

    Leonard F. Pease III

  5. Department of Pharmaceutics & Pharmaceutical Chemistry, University of Utah, Salt Lake City, UT, 84112, USA

    Leonard F. Pease III

  6. Department of Pathology, University of Utah, North Medical Drive East, Salt Lake City, UT, 84112, USA

    Philip S. Bernard

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  1. Vasiliy S. Chernyshev
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  2. Rakesh Rachamadugu
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Correspondence to Mikhail Skliar.

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Chernyshev, V.S., Rachamadugu, R., Tseng, Y.H. et al. Size and shape characterization of hydrated and desiccated exosomes. Anal Bioanal Chem 407, 3285–3301 (2015). https://doi.org/10.1007/s00216-015-8535-3

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