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. 2016 Feb 25:6:21933.
doi: 10.1038/srep21933.

Engineering hybrid exosomes by membrane fusion with liposomes

Yuko T Sato  1 , Kaori Umezaki  1 , Shinichi Sawada  1   2 , Sada-atsu Mukai  1   2 , Yoshihiro Sasaki  2 , Naozumi Harada  3 , Hiroshi Shiku  3 , Kazunari Akiyoshi  1   2
Affiliations

Engineering hybrid exosomes by membrane fusion with liposomes

Yuko T Sato et al. Sci Rep. .

Abstract

Exosomes are a valuable biomaterial for the development of novel nanocarriers as functionally advanced drug delivery systems. To control and modify the performance of exosomal nanocarriers, we developed hybrid exosomes by fusing their membranes with liposomes using the freeze-thaw method. Exosomes embedded with a specific membrane protein isolated from genetically modified cells were fused with various liposomes, confirming that membrane engineering methods can be combined with genetic modification techniques. Cellular uptake studies performed using the hybrid exosomes revealed that the interactions between the developed exosomes and cells could be modified by changing the lipid composition or the properties of the exogenous lipids. These results suggest that the membrane-engineering approach reported here offers a new strategy for developing rationally designed exosomes as hybrid nanocarriers for use in advanced drug delivery systems.

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Figures

Figure 1
Figure 1. Schematic of method used to engineer the exosome–liposome hybrids.
Figure 2
Figure 2. Characterization of Raw 264.7 exosomes.
(a) Western blot analysis of exosomes using anti-CD9 or anti-Hsc70 antibodies. (b) Size distribution profile, as determined by nanoparticle tracking analysis.
Figure 3
Figure 3. Interaction of Raw 264.7 exosomes with liposomes containing various lipids.
(a) Fluorescence spectra with excitation at 460 nm. The light grey, dark gray and black lines represent the spectra before, 5 times and 10 times after freeze thawing, respectively. (b) Relationship between the number of freeze–thaw cycles and the dilution ratios of DOPC liposomes or exosome–DOPC liposomes (left), the dilution ratios of DOPS liposomes or exosome–DOPS (right).
Figure 4
Figure 4. Characterization of CMS7 exosomes.
(a,b) FACS analysis of CD63 expression on the surface of exosomes from CMS7-wt (a) and CMS7-HE (b) cells. The grey and black lines represent isotype controls and anti-CD63, respectively. (c,d) Size distribution profile, as determined by nanoparticle tracking analysis, of exosomes from CMS7-wt (c) and CMS7-HE (d) cells. (e–g) Western blot analysis of exosomes from CMS7-wt or CMS7-HE cells using anti-Hsc70 (e), anti-HER2 (f), or anti-phosphorylated HER2 (g) antibodies.
Figure 5
Figure 5. Western blot analysis of CMS7-HE exosomes following membrane fusion with DOTAP liposomes.
Western blotting was conducted using anti-HER2 (a) or anti-phosphorylated HER2 (b) antibodies.
Figure 6
Figure 6. Cellular uptake of exosomes or exosome–liposome hybrids.
(a) Mean fluorescence of cells, as measured by flow cytometry. (b) Confocal laser scanning microscopy images of HeLa cells incubated with exosomes or exosome–liposome hybrid. Green fluorescence represents 5- or 6-(N-succinimidyloxycarbonyl)-fluorescein 3′,6′-diacetate-labelled exosome.
See this image and copyright information in PMC

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References

    1. Valadi H. et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat. Cell Biol. 9, 654–659 (2007). - PubMed
    1. Properzi F., Logozzi M. & Fais S. Exosomes: the future of biomarkers in medicine. Biomark. Med. 7, 769–778 (2013). - PubMed
    1. Jia S. et al. Emerging technologies in extracellular vesicle-based molecular diagnostics. Expert Rev. Mol. Diagn. 14, 307–321 (2014). - PubMed
    1. Mulcahy L. A., Pink R. C. & Carter D. R. F. Routes and mechanisms of extracellular vesicle uptake. J. Extracell. Vesicles 3, 1–14 (2014). - PMC - PubMed
    1. Parolini I. et al. Microenvironmental pH is a key factor for exosome traffic in tumor cells. J. Biol. Chem. 284, 34211–34222 (2009). - PMC - PubMed

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