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. 2018 Jun 8;14(1):179.
doi: 10.1186/s12917-018-1509-x.

Exploration of serum- and cell culture-derived exosomes from dogs

Matias Aguilera-Rojas  1 , Brit Badewien-Rentzsch  1 , Johanna Plendl  2 , Barbara Kohn  3 , Ralf Einspanier  4
Affiliations

Exploration of serum- and cell culture-derived exosomes from dogs

Matias Aguilera-Rojas et al. BMC Vet Res. .

Abstract

Background: Exosomes are defined as extracellular membrane vesicles, 30-150 nm in diameter, derived from all types of cells. They originate via endocytosis and then they are released through exocytosis to the extracellular space, being found in various biological fluids as well as in cell culture medium. In the last few years, exosomes have gained considerable scientific interest due to their potential use as biomarkers, especially in the field of cancer research. This report describes a method to isolate, quantify and identify serum- and cell culture-derived exosomes from dog samples, using small volumes (100 μL and 1 mL, respectively).

Results: Quantification and sizing of exosomes contained in serum and cell culture samples were assessed by utilizing nanoparticle tracking analysis, transmission electron microscopy and immunoelectron microscopy. Detected particles showed the normal size (30-150 nm) and morphology described for exosomes, as well as presence of the transmembrane protein CD63 known as exosomal marker.

Conclusions: Based on a validated rapid isolation procedure of nanoparticles from small volumes of different types of dog samples, a characterization and exploration of intact exosomes, as well as facilitation for their analysis in downstream applications was introduced.

Keywords: Biomarkers; Cell culture medium; Dog; Exosomes; Nanoparticle tracking analysis; Serum; Transmission electron microscopy.

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Conflict of interest statement

Ethics approval and consent to participate

The ethics committee from the Governmental Office for Health and Social Affairs Berlin (LAGeSo Berlin), file number StN 0005/17, do not classify the use of residual blood from diagnostic samples from dogs or cats and the use of euthanized animals as animal experimentation.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Nano track analysis size distribution of exosomes isolated from samples of canine origin. a blood serum-derived exosomes, b C2 cell line culture-derived exosomes and (c) primary fibroblasts culture-derived exosomes. Red error bars indicate +/− standard error of the mean
Fig. 2
Fig. 2
Transmission electron microscopy of native exosomes isolated from samples of canine origin. a blood serum-derived exosomes, b C2 cell line culture-derived exosomes and (c and d) primary fibroblasts culture-derived exosomes. Size bar = 100 nm
Fig. 3
Fig. 3
Immunoelectron microscopy images of exosomes isolated from samples of canine origin. a and b serum-derived exosomes, c C2 cell line culture-derived exosomes, d and e primary fibroblasts culture-derived exosomes. Note the gold particles bound to the exosome membrane indicating presence of the tetraspanin CD63. Size bar = 100 nm
See this image and copyright information in PMC

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