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Production of Extracellular Vesicles Loaded with Therapeutic Cargo

  • Tek N. Lamichhane
  • Steven M. JayEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1831)

Abstract

Extracellular vesicles (EVs) are biological nanoparticles comprising exosomes, microvesicles, and other heterogeneous nanoscopic vesicle populations that are produced by most cell types. In addition to their putative roles as critical mediators of intercellular communication, EVs have begun to be harnessed as drug delivery vehicles, with early evidence indicating they may have significant advantages over synthetic nanoparticle delivery systems for particular applications. Targeted delivery of EV-encapsulated cargo has already been realized and may have broad applicability; however, methods for producing and purifying EVs and loading them with therapeutic molecules have yet to be standardized. In this chapter, we outline steps for EV isolation and characterization and compare current methods for active and passive loading of EVs with payloads of short interfering RNA (siRNA) or small molecules, with the results revealing that active loading via electroporation increases loading efficiency of siRNA but not of Rhodamine B, a model for a small molecule drug, in HEK293T-derived EVs. The methods described here may inform future design of targeted delivery of nucleic acids or small molecules via EVs.

Key words

Extracellular vesicles (EVs) Exosomes siRNA Small molecules Electroporation Drug delivery Cancer therapeutics 

Notes

Acknowledgments

This work was supported by NIH R00 grant HL112905, by an ORAU Ralph E. Power Junior Faculty Enhancement Award, and by two University of Maryland Tier 1 seed grants (all to S.M.J.). The authors thank Rini Pek, Navein Arumugasaamy and Anjana Jeyaram for their helpful contributions.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Fischell Department of BioengineeringUniversity of MarylandCollege ParkUSA
  2. 2.Program in Oncology, Marlene and Stewart Greenebaum Cancer CenterUniversity of MarylandCollege ParkUSA
  3. 3.Program in Molecular and Cell BiologyUniversity of MarylandCollege ParkUSA

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