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Isolation of Human Small Extracellular Vesicles and Tracking of Their Uptake by Retinal Pigment Epithelial Cells In Vitro

1
Division of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, Switzerland
2
Institute of Parasitology, Vetsuisse Faculty, University of Bern, 3012 Bern, Switzerland
*
Author to whom correspondence should be addressed.
Co-first authors.
Int. J. Mol. Sci. 2020, 21(11), 3799; https://doi.org/10.3390/ijms21113799
Received: 1 April 2020 / Revised: 21 May 2020 / Accepted: 24 May 2020 / Published: 27 May 2020
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
Small extracellular vesicles (EVs) are among the most frequently investigated EVs and play major roles in intercellular communication by delivering various cargo molecules to target cells. They could potentially represent an alternative delivery strategy to treat ocular toxoplasmosis, a parasitosis affecting the retinal pigment epithelium (RPE). To date, the uptake of human small EVs by RPE cells has never been reported. In this study, we report on the intracellular uptake of fluorescently labelled human urine and fibroblast-derived small EVs by human RPE cells. In summary, both dye-labelled urinary small EVs and small EVs obtained from fibroblasts stably expressing membrane-bound green fluorescent protein were successfully internalized by RPE cells as revealed by immunohistochemistry. In recipient ARPE19 cells, BODIPY-labelled small EVs were found in close vicinity to the parasite Toxoplasma gondii. Additionally, an ultrastructural method was enabled to distinguish between labelled exogenous and endogenous small EVs within target cells. View Full-Text
Keywords: small extracellular vesicles; BODIPY; PKH; EV; retinal pigment epithelium; T. gondii small extracellular vesicles; BODIPY; PKH; EV; retinal pigment epithelium; T. gondii
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MDPI and ACS Style

Marcu, I.C.; Eberhard, N.; Yerly, A.; Balmer, V.; Hemphill, A.; Mogel, H.; Gaschen, V.; Stoffel, M.H.; Bluteau, J. Isolation of Human Small Extracellular Vesicles and Tracking of Their Uptake by Retinal Pigment Epithelial Cells In Vitro. Int. J. Mol. Sci. 2020, 21, 3799. https://doi.org/10.3390/ijms21113799

AMA Style

Marcu IC, Eberhard N, Yerly A, Balmer V, Hemphill A, Mogel H, Gaschen V, Stoffel MH, Bluteau J. Isolation of Human Small Extracellular Vesicles and Tracking of Their Uptake by Retinal Pigment Epithelial Cells In Vitro. International Journal of Molecular Sciences. 2020; 21(11):3799. https://doi.org/10.3390/ijms21113799

Chicago/Turabian Style

Marcu, Irene C., Naja Eberhard, Anaïs Yerly, Verena Balmer, Andrew Hemphill, Helga Mogel, Véronique Gaschen, Michael H. Stoffel, and Jasmin Bluteau. 2020. "Isolation of Human Small Extracellular Vesicles and Tracking of Their Uptake by Retinal Pigment Epithelial Cells In Vitro" International Journal of Molecular Sciences 21, no. 11: 3799. https://doi.org/10.3390/ijms21113799

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