Abstract
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.
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Abbreviations
- AFM:
-
Atomic force microscopy
- ASC:
-
Adipose-derived stem cells
- BBB:
-
Blood–brain barrier
- CD:
-
Cluster of differentiation
- CFSE:
-
Carboxyfluorescein succinimidyl ester
- CM:
-
Conditioned medium
- EE:
-
Electroporated exosomes
- ESCRT:
-
Endosomal sorting complex required for transport
- FCS:
-
Fetal calf serum
- GBM:
-
Glioblastoma multiforme
- miRNA:
-
MicroRNA
- mRNA:
-
Messenger RNA
- MSC:
-
Mesenchymal stem cell
- MVB:
-
Multivesicular bodies
- NTA:
-
Nanoparticle tracking analysis
- PEG:
-
Polyethylene glycol
- Pen/strep:
-
Penicillin/streptomycin
- PTA:
-
Phosphotungstic acid
- RNA:
-
Ribonucleic acid
- siRNA:
-
Small interfering RNA
- TEM:
-
Transmission electron microscopy
- TPM:
-
Trehalose pulse medium
- UC:
-
Ultracentrifugation
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Acknowledgments
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).
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Johnsen, K.B., Gudbergsson, J.M., Skov, M.N. et al. Evaluation of electroporation-induced adverse effects on adipose-derived stem cell exosomes. Cytotechnology 68, 2125–2138 (2016). https://doi.org/10.1007/s10616-016-9952-7
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DOI: https://doi.org/10.1007/s10616-016-9952-7