Abstract
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.
Highlights
Extracellular vesicles (EVs) are membrane-surrounded vesicles released into the extracellular space by cells
Labelling of EVs Using Lactadherin Conjugated to a Fluorescent Tag
The results showed that labeled EVs were removed unbound lactadherin from EVs
Summary
Extracellular vesicles (EVs) are membrane-surrounded vesicles released into the extracellular space by cells. EVs have been recognised as important vehicles for intercellular communication. This kind of vesicle has been isolated from many different tissues, healthy and diseased, and every biological fluid in the body, including saliva, blood, urine, and milk. The soluble core of EVs contains mRNA and microRNA (miRNA) with the potential to modulate the recipient cell function [1]. These shed membrane vesicles are suggested to have various biological functions and potentially as natural carriers for drug delivery [2]
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