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Article

Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells

1
Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, Denmark
2
Department of Clinical Medicine, Aarhus University, 8000 Aarhus, Denmark
*
Authors to whom correspondence should be addressed.
Present affiliation: Department of Chemical Engineering, Biotechnology and Environmental Technology, University of Southern Denmark, 5230 Odense, Denmark.
Biomedicines 2020, 8(7), 211; https://doi.org/10.3390/biomedicines8070211
Received: 26 June 2020 / Revised: 10 July 2020 / Accepted: 10 July 2020 / Published: 14 July 2020
The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacity to modulate the infant’s intestinal and immune system. Studies suggest that milk EVs are enriched in immune-modulating proteins and miRNA, highlighting their possible health benefits to infants. To assess uptake of milk EVs by intestinal epithelial cells, a method was developed using labelling of isolated EVs with fluorophore-conjugated lactadherin. Lactadherin is a generic and validated EV marker, which enables an effective labelling of phosphatidylserine (PS) exposing EVs. Labelled EVs could effectively be used to describe a dose- and time-dependent uptake into the intestinal epithelial Caco-2 cell line. Additionally, fluorescence microscopy was employed to show that EVs colocalize with endosomal markers and lysosomes, indicating that EVs are taken up via general endocytotic mechanisms. Collectively, a method to specifically label isolated EVs is presented and employed to study the uptake of milk EVs by intestinal epithelial cells. View Full-Text
Keywords: milk; extracellular vesicles; exosomes; labelling; lactadherin; fluorescence; uptake; intestinal epithelium; Caco-2; endosomes milk; extracellular vesicles; exosomes; labelling; lactadherin; fluorescence; uptake; intestinal epithelium; Caco-2; endosomes
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MDPI and ACS Style

Hansen, M.S.; Gadegaard, I.S.E.; Arnspang, E.C.; Blans, K.; Nejsum, L.N.; Rasmussen, J.T. Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells. Biomedicines 2020, 8, 211. https://doi.org/10.3390/biomedicines8070211

AMA Style

Hansen MS, Gadegaard ISE, Arnspang EC, Blans K, Nejsum LN, Rasmussen JT. Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells. Biomedicines. 2020; 8(7):211. https://doi.org/10.3390/biomedicines8070211

Chicago/Turabian Style

Hansen, Maria S., Ida S.E. Gadegaard, Eva C. Arnspang, Kristine Blans, Lene N. Nejsum, and Jan T. Rasmussen 2020. "Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells" Biomedicines 8, no. 7: 211. https://doi.org/10.3390/biomedicines8070211

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