title: In vitro toxicology studies of extracellular vesicles.
abstract: Extracellular vesicles (EVs) are membrane-bound vesicles released from cells into the extracellular environment. There is emerging interest in the use of EVs as potential therapeutic interventions. We sought to evaluate the safety of EVs that may be therapeutically used by performing in vitro toxicological assessments. EVs were obtained from mesenchymal stem cells (MSC-EV) or from bovine milk (BM-EV) by differential ultracentrifugation, and quantitated using nanoparticle tracking analysis. Genotoxic effects, hematological effects, immunological effects and endotoxin production were evaluated at two dose levels. Neither MSC-EVs nor BM-EVs elicited detectable genotoxic effects using either the alkaline comet assay or micronucleus assay. Hemolysis was observed with BM-EVs but not with MSC-EVs. MSC-EVs did not have any significant effect on either spontaneous or collagen-induced platelet aggregation. In contrast, BM-EVs were noted to increase collagen-induced platelet aggregation, even though no spontaneous increase in platelet aggregation was noted. Both types of EVs induced leukocyte proliferation, which was greater with BM-EV. Neither MSC-EVs nor BM-EVs induced HL-60 phagocytosis, although BM-EVs decreased zymosan-induced phagocytosis. Furthermore, neither MSC-EVs nor BM-EVs induced nitric oxide production. Unlike MSC-EVs, BM-EVs tested positive for endotoxin and induced complement activation. There are significant differences in toxicological profiles between MSC-EVs and BM-EVs that may reflect variations in techniques for EV isolation, EV content or cross-species differences. The safety of MSC-EV supports their use for disease therapeutics, whereas detailed safety and toxicological assessment will be necessary before the use of BM-EVs. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Department of Transplantation and Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Sayantan
authors.identifier: 0000-0002-3494-8129
authors.initials: S
authors.lastname: Maji
authors.affiliation: Department of Transplantation and Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Irene K
authors.initials: IK
authors.lastname: Yan
authors.affiliation: Department of Transplantation and Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Mansi
authors.initials: M
authors.lastname: Parasramka
authors.affiliation: Department of Transplantation and Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Swathi
authors.initials: S
authors.lastname: Mohankumar
authors.affiliation: Department of Transplantation and Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Akiko
authors.initials: A
authors.lastname: Matsuda
authors.affiliation: Department of Transplantation and Department of Cancer Biology, Mayo Clinic, Jacksonville, FL, USA.
authors.forename: Tushar
authors.initials: T
authors.lastname: Patel
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D003412:Cricetulus; D064420:Drug-Related Side Effects and Adverse Reactions; D000067128:Extracellular Vesicles / Q000737:chemistry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D051379:Mice; D008892:Milk / Q000737:chemistry; D018675:Toxicity Tests / Q000379:methods*
keywords: bovine milk-extracellular vesicles; extracellular vesicles; mesenchymal stem cell-extracellular vesicles; toxicity