In the recent years, the possibility of utilizing extracellular vesicles for drug delivery purposes has been investigated in various models, suggesting that these vesicles may have such potential. In addition to the choice of donor cell type for vesicle production, a major obstacle still exists with respect of loading the extracellular vesicles efficiently with the drug of choice. One of the proposed solutions to this problem has been drug loading by electroporation, where small pores are created in the membrane of the extracellular vesicles, hereby allowing for free diffusion of the drug compound into the interior of the vesicle. We investigated the utility of adipose-derived stem cells (ASCs) as an efficient exosome donor cell type with a particular focus on the treatment of glioblastoma multiforme (GBM). In addition, we evaluated electroporation-induced effects on the ASC exosomes with respect to their endogenous potential of stimulating GBM proliferation, and morphological changes to single and multiple ASC exosomes. We found that electroporation does not change the endogenous stimulatory capacity of ASC exosomes on GBM cell proliferation, but mediates adverse morphological changes including aggregation of the exosomes. In order to address this issue, we have successfully optimized the use of a trehalose-containing buffer system as a way of maintaining the structural integrity of the exosomes.
This is a preview of subscription content, log in to check access.
The authors would like to acknowledge laboratory technician Rikke Sophie Holm Kristensen, Aalborg University for her excellent technical assistance. Furthermore, Andreas Rasmussen, Laboratory of Stem Cell Research, Aalborg University is acknowledged for his kind help and facilitation of electroporation. This work was supported by Spar Nord Fonden. Kasper Bendix Johnsen is supported by the Novo Scholarship Programme (Novo Nordisk, Denmark).
Chen TS, Arslan F, Yin Y et al (2011) Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs. J Transl Med 9:47. doi:10.1186/1479-5876-9-47CrossRefGoogle Scholar
Crowe JH, Crowe LM (1988) Factors affecting the stability of dry liposomes. Biochim Biophys Acta 939:327–334CrossRefGoogle Scholar
de Jong OG, Verhaar MC, Chen Y et al (2012) Cellular stress conditions are reflected in the protein and RNA content of endothelial cell-derived exosomes. J Extracell Vesicles 1:569. doi:10.3402/jev.v1i0.18396Google Scholar
El Andaloussi SE, Mäger I, Breakefield XO, Wood MJA (2013) Extracellular vesicles: biology and emerging therapeutic opportunities. Nat Rev Drug Discov. doi:10.1038/nrd3978Google Scholar
Johnsen KB, Gudbergsson JM, Skov MN et al (2014) A comprehensive overview of exosomes as drug delivery vehicles—endogenous nanocarriers for targeted cancer therapy. Biochim Biophys Acta. doi:10.1016/j.bbcan.2014.04.005Google Scholar
Kapla J, Engström O, Stevensson B et al (2015) Molecular dynamics simulations and NMR spectroscopy studies of trehalose–lipid bilayer systems. Phys Chem Chem Phys 17:22438–22447. doi:10.1039/c5cp02472bCrossRefGoogle Scholar
Kooijmans SAA, Stremersch S, Braeckmans K et al (2013) Electroporation-induced siRNA precipitation obscures the efficiency of siRNA loading into extracellular vesicles. J Control Release. doi:10.1016/j.jconrel.2013.08.014Google Scholar
Koster KL, Webb MS, Bryant G, Lynch DV (1994) Interactions between soluble sugars and POPC (1-palmitoyl-2-oleoylphosphatidylcholine) during dehydration: vitrification of sugars alters the phase behavior of the phospholipid. Biochim Biophys Acta 1193:143–150CrossRefGoogle Scholar
Lee HK, Finniss S, Cazacu S et al (2013) Mesenchymal stem cells deliver synthetic microRNA mimics to glioma cells and glioma stem cells and inhibit their cell migration and self-renewal. Oncotarget 4:346–361CrossRefGoogle Scholar
Lin R, Wang S, Zhao RC (2013) Exosomes from human adipose-derived mesenchymal stem cells promote migration through Wnt signaling pathway in a breast cancer cell model. Mol Cell Biochem. doi:10.1007/s11010-013-1746-zGoogle Scholar
Lopatina T, Bruno S, Tetta C et al (2014) Platelet-derived growth factor regulates the secretion of extracellular vesicles by adipose mesenchymal stem cells and enhance their angiogenic potential. Cell Commun Signal 12:26. doi:10.1186/1478-811X-12-26CrossRefGoogle Scholar
Pereira CS, Hünenberger PH (2006) Interaction of the sugars trehalose, maltose and glucose with a phospholipid bilayer: a comparative molecular dynamics study. J Phys Chem B 110:15572–15581. doi:10.1021/jp060789lCrossRefGoogle Scholar
Rasmussen JG, Frøbert O, Pilgaard L et al (2011) Prolonged hypoxic culture and trypsinization increase the pro-angiogenic potential of human adipose tissue-derived stem cells. Cytotherapy 13:318–328. doi:10.3109/14653249.2010.506505CrossRefGoogle Scholar
Villarreal MA, Díaz SB, Disalvo EA, Montich GG (2004) Molecular dynamics simulation study of the interaction of trehalose with lipid membranes. Langmuir 20:7844–7851. doi:10.1021/la049485lCrossRefGoogle Scholar
Vlassov AV, Magdaleno S, Setterquist R, Conrad R (2012) Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim Biophys Acta 1820:940–948. doi:10.1016/j.bbagen.2012.03.017CrossRefGoogle Scholar
Wahlgren J, Karlson TDL, Brisslert M et al (2012) Plasma exosomes can deliver exogenous short interfering RNA to monocytes and lymphocytes. Nucleic Acids Res. doi:10.1093/nar/gks463Google Scholar
Weaver JC (1993) Electroporation: a general phenomenon for manipulating cells and tissues. J Cell Biochem 51:426–435CrossRefGoogle Scholar
Witwer KW, Buzás EI, Bemis LT et al (2013) Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J Extracell Vesicles 2:18389. doi:10.1074/jbc.M111.277061Google Scholar
Yang S, Pilgaard L, Chase LG et al (2012) Defined xenogeneic-free and hypoxic environment provides superior conditions for long-term expansion of human adipose-derived stem cells. Tissue Eng Part C Methods 18:593–602. doi:10.1089/ten.TEC.2011.0592CrossRefGoogle Scholar