Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro
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...
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doaj-a35e1c5ed3864d94b6f22794f52becad2020-11-25T03:17:20ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-05-01213799379910.3390/ijms21113799Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In VitroIrene C. Marcu0Naja Eberhard1Anaïs Yerly2Verena Balmer3Andrew Hemphill4Helga Mogel5Véronique Gaschen6Michael H. Stoffel7Jasmin Bluteau8Division of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandInstitute of Parasitology, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandDivision of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandInstitute of Parasitology, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandInstitute of Parasitology, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandDivision of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandDivision of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandDivision of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandDivision of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, 3012 Bern, SwitzerlandSmall 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 <i>Toxoplasma gondii.</i> Additionally, an ultrastructural method was enabled to distinguish between labelled exogenous and endogenous small EVs within target cells.https://www.mdpi.com/1422-0067/21/11/3799small extracellular vesiclesBODIPYPKHEVretinal pigment epithelium<i>T. gondii</i> |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Irene C. Marcu Naja Eberhard Anaïs Yerly Verena Balmer Andrew Hemphill Helga Mogel Véronique Gaschen Michael H. Stoffel Jasmin Bluteau |
spellingShingle |
Irene C. Marcu Naja Eberhard Anaïs Yerly Verena Balmer Andrew Hemphill Helga Mogel Véronique Gaschen Michael H. Stoffel Jasmin Bluteau Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro International Journal of Molecular Sciences small extracellular vesicles BODIPY PKH EV retinal pigment epithelium <i>T. gondii</i> |
author_facet |
Irene C. Marcu Naja Eberhard Anaïs Yerly Verena Balmer Andrew Hemphill Helga Mogel Véronique Gaschen Michael H. Stoffel Jasmin Bluteau |
author_sort |
Irene C. Marcu |
title |
Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro |
title_short |
Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro |
title_full |
Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro |
title_fullStr |
Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro |
title_full_unstemmed |
Isolation of Human Small Extracellular Vesicles and Tracking of their Uptake by Retinal Pigment Epithelial Cells In Vitro |
title_sort |
isolation of human small extracellular vesicles and tracking of their uptake by retinal pigment epithelial cells in vitro |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2020-05-01 |
description |
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 <i>Toxoplasma gondii.</i> Additionally, an ultrastructural method was enabled to distinguish between labelled exogenous and endogenous small EVs within target cells. |
topic |
small extracellular vesicles BODIPY PKH EV retinal pigment epithelium <i>T. gondii</i> |
url |
https://www.mdpi.com/1422-0067/21/11/3799 |
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