title: Maximizing exosome colloidal stability following electroporation.
abstract: 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.
authors: Consortium for Translational Research in Advanced Imaging and Nanomedicine (C-TRAIN), Cardiovascular Division, Department of Medicine, Washington University School of Medicine, 4320 Forest Park Avenue, Suite 101, Campus Box 8215, St. Louis, MO 63108, USA. Electronic address: jhood@dom.wustl.edu.
authors: Joshua L
authors: JL
authors: Hood
authors: Consortium for Translational Research in Advanced Imaging and Nanomedicine (C-TRAIN), Cardiovascular Division, Department of Medicine, Washington University School of Medicine, 4320 Forest Park Avenue, Suite 101, Campus Box 8215, St. Louis, MO 63108, USA. Electronic address: mike@dom.wustl.edu.
authors: Michael J
authors: MJ
authors: Scott
authors: Consortium for Translational Research in Advanced Imaging and Nanomedicine (C-TRAIN), Cardiovascular Division, Department of Medicine, Washington University School of Medicine, 4320 Forest Park Avenue, Suite 101, Campus Box 8215, St. Louis, MO 63108, USA. Electronic address: Wicklines@aol.com.
authors: Samuel A
authors: SA
authors: Wickline
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003102:Colloids / Q000737:chemistry*; D018274:Electroporation; D055354:Exosomes / Q000737:chemistry / Q000378:metabolism*; D052203:Ferrosoferric Oxide / Q000737:chemistry; D006801:Humans; D008027:Light; D058185:Magnetite Nanoparticles / Q000737:chemistry; D051379:Mice; D010316:Particle Size; D012313:RNA / Q000378:metabolism; D012542:Scattering, Radiation; D014199:Trehalose / Q000737:chemistry
chemical_list: D003102:Colloids; D058185:Magnetite Nanoparticles; D012313:RNA; D014199:Trehalose; D052203:Ferrosoferric Oxide
keywords: Electroporation; Exosomes; Iron oxide; Superparamagnetic; Trehalose