Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells
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...
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doaj-fd8bcfe881b94d6e9c363921ce80274b2020-11-25T03:21:34ZengMDPI AGBiomedicines2227-90592020-07-01821121110.3390/biomedicines8070211Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial CellsMaria S. Hansen0Ida S. E. Gadegaard1Eva C. Arnspang2Kristine Blans3Lene N. Nejsum4Jan T. Rasmussen5Department of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, DenmarkDepartment of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8000 Aarhus, DenmarkDepartment of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, DenmarkDepartment of Clinical Medicine, Aarhus University, 8000 Aarhus, DenmarkDepartment of Molecular Biology and Genetics, Aarhus University, 8000 Aarhus, DenmarkThe 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.https://www.mdpi.com/2227-9059/8/7/211milkextracellular vesiclesexosomeslabellinglactadherinfluorescence |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Maria S. Hansen Ida S. E. Gadegaard Eva C. Arnspang Kristine Blans Lene N. Nejsum Jan T. Rasmussen |
spellingShingle |
Maria S. Hansen Ida S. E. Gadegaard Eva C. Arnspang Kristine Blans Lene N. Nejsum Jan T. Rasmussen Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells Biomedicines milk extracellular vesicles exosomes labelling lactadherin fluorescence |
author_facet |
Maria S. Hansen Ida S. E. Gadegaard Eva C. Arnspang Kristine Blans Lene N. Nejsum Jan T. Rasmussen |
author_sort |
Maria S. Hansen |
title |
Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells |
title_short |
Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells |
title_full |
Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells |
title_fullStr |
Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells |
title_full_unstemmed |
Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells |
title_sort |
specific and non-invasive fluorescent labelling of extracellular vesicles for evaluation of intracellular processing by intestinal epithelial cells |
publisher |
MDPI AG |
series |
Biomedicines |
issn |
2227-9059 |
publishDate |
2020-07-01 |
description |
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. |
topic |
milk extracellular vesicles exosomes labelling lactadherin fluorescence |
url |
https://www.mdpi.com/2227-9059/8/7/211 |
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