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

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Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells. / Hansen, Maria Stenum; Gadegaard, Ida Schnack Eg; Christensen, Eva Arnspang; Blans, Kristine Ingrid Marie; Nejsum, Lene Niemann; Rasmussen, Jan Trige.

In: Biomedicines, Vol. 8, No. 7, 211, 2020.

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@article{4084d0f17ea142b8b538bd21c39b862e,
title = "Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells",
abstract = "The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacity to modulate the infant{\textquoteright}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.",
author = "Hansen, {Maria Stenum} and Gadegaard, {Ida Schnack Eg} and Christensen, {Eva Arnspang} and Blans, {Kristine Ingrid Marie} and Nejsum, {Lene Niemann} and Rasmussen, {Jan Trige}",
year = "2020",
doi = "10.3390/biomedicines8070211",
language = "English",
volume = "8",
journal = "Biomedicines",
issn = "2227-9059",
publisher = "MDPI",
number = "7",

}

RIS

TY - JOUR

T1 - Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells

AU - Hansen, Maria Stenum

AU - Gadegaard, Ida Schnack Eg

AU - Christensen, Eva Arnspang

AU - Blans, Kristine Ingrid Marie

AU - Nejsum, Lene Niemann

AU - Rasmussen, Jan Trige

PY - 2020

Y1 - 2020

N2 - 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.

AB - 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.

U2 - 10.3390/biomedicines8070211

DO - 10.3390/biomedicines8070211

M3 - Journal article

C2 - 32674302

VL - 8

JO - Biomedicines

JF - Biomedicines

SN - 2227-9059

IS - 7

M1 - 211

ER -