Maximizing exosome colloidal stability following electroporation

Development of exosome-based semisynthetic nanovesicles for diagnostic and therapeutic purposes requires novel approaches to load exosomes with cargo. Electroporation has previously been used to load exosomes with RNA. However, investigations into exosome colloidal stability following electroporatio...

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Bibliographic Details
Published in:Analytical biochemistry 2014-03, Vol.448, p.41-49
Main Authors: Hood, Joshua L., Scott, Michael J., Wickline, Samuel A.
Format: Article
Language:English
Subjects:
Animals
Cell Line, Tumor
Colloids - chemistry
Electroporation
Exosomes
Exosomes - chemistry
Exosomes - metabolism
Ferrosoferric Oxide - chemistry
Humans
Iron oxide
Light
Magnetite Nanoparticles - chemistry
Mice
Particle Size
RNA - metabolism
Scattering, Radiation
Superparamagnetic
Trehalose
Trehalose - chemistry
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Summary:Development of exosome-based semisynthetic nanovesicles for diagnostic and therapeutic purposes requires novel approaches to load exosomes with cargo. Electroporation has previously been used to load exosomes with RNA. However, investigations into exosome colloidal stability following electroporation have not been considered. Herein, we report the development of a unique trehalose pulse media (TPM) that minimizes exosome aggregation following electroporation. Dynamic light scattering (DLS) and RNA absorbance were employed to determine the extent of exosome aggregation and electroextraction post electroporation in TPM compared to common PBS pulse media or sucrose pulse media (SPM). Use of TPM to disaggregate melanoma exosomes post electroporation was dependent on both exosome concentration and electric field strength. TPM maximized exosome dispersal post electroporation for both homogenous B16 melanoma and heterogeneous human serum-derived populations of exosomes. Moreover, TPM enabled heavy cargo loading of melanoma exosomes with 5nm superparamagnetic iron oxide nanoparticles (SPION5) while maintaining original exosome size and minimizing exosome aggregation as evidenced by transmission electron microscopy. Loading exosomes with SPION5 increased exosome density on sucrose gradients. This provides a simple, label-free means of enriching exogenously modified exosomes and introduces the potential for MRI-driven theranostic exosome investigations in vivo.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2013.12.001