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Evaluation of electroporation-induced adverse effects on adipose-derived stem cell exosomes

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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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Authors and Affiliations

  1. Laboratory for Cancer Biology, Institute of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg Ø, Denmark

    Kasper Bendix Johnsen, Johann Mar Gudbergsson, Martin Najbjerg Skov & Meg Duroux

  2. Laboratory for Neurobiology, Institute of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 3B, 9220, Aalborg Ø, Denmark

    Kasper Bendix Johnsen & Torben Moos

  3. Department of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, Aarhus C, 8000, Aarhus, Denmark

    Gunna Christiansen

  4. Institute of Physics and Nanotechnology, Aalborg University, Skjernvej 4, 9220, Aalborg Ø, Denmark

    Leonid Gurevich

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  1. Kasper Bendix Johnsen
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  2. Johann Mar Gudbergsson
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Correspondence to Meg Duroux.

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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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