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 S. E.
AU - Arnspang, Eva C.
AU - Blans, Kristine
AU - Nejsum, Lene
AU - Rasmussen, Jan
PY - 2020/7/14
Y1 - 2020/7/14
N2 - The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacityto modulate the infant’s intestinal and immune system. Studies suggest that milk EVs are enrichedin 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 ofisolated EVs with fluorophore-conjugated lactadherin. Lactadherin is a generic and validated EVmarker, which enables an effective labelling of phosphatidylserine (PS) exposing EVs. Labelled EVscould effectively be used to describe a dose- and time-dependent uptake into the intestinal epithelialCaco-2 cell line. Additionally, fluorescence microscopy was employed to show that EVs colocalizewith endosomal markers and lysosomes, indicating that EVs are taken up via general endocytoticmechanisms. Collectively, a method to specifically label isolated EVs is presented and employed tostudy the uptake of milk EVs by intestinal epithelial cells.
AB - The presence of extracellular vesicles (EVs) in milk has gained interest due to their capacityto modulate the infant’s intestinal and immune system. Studies suggest that milk EVs are enrichedin 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 ofisolated EVs with fluorophore-conjugated lactadherin. Lactadherin is a generic and validated EVmarker, which enables an effective labelling of phosphatidylserine (PS) exposing EVs. Labelled EVscould effectively be used to describe a dose- and time-dependent uptake into the intestinal epithelialCaco-2 cell line. Additionally, fluorescence microscopy was employed to show that EVs colocalizewith endosomal markers and lysosomes, indicating that EVs are taken up via general endocytoticmechanisms. Collectively, a method to specifically label isolated EVs is presented and employed tostudy 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
ER -