Bioavailability of bovine milk extracellular vesicles

Background: Milk extracellular vesicles (MEVs) are a novel class of milk bioactives, which most likely are resistant to the digestive system after consumption. Around the world, people drink milk from many animals such as cow, camel, goat and sheep, with cow's milk being the most consumed type....

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Bibliographic Details
Published inJournal of extracellular vesicles Vol. 7; p. 218
Main Authors Hansen, Maria S, Blans, Kristine I M, Rasmussen, Jan T
Format Journal Article
LanguageEnglish
Published Abingdon John Wiley & Sons, Inc 01.01.2018
Subjects
Bioavailability
Biological activity
Casein
Cell lines
Centrifugation
Contaminants
Cow's milk
Epithelial cells
Epithelium
Extracellular vesicles
Internalization
Intestine
Investigations
Labeling
Milk
Milk fat globule membranes
Denmark
Aarhus Denmark
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Summary:Background: Milk extracellular vesicles (MEVs) are a novel class of milk bioactives, which most likely are resistant to the digestive system after consumption. Around the world, people drink milk from many animals such as cow, camel, goat and sheep, with cow's milk being the most consumed type. As bovine milk is a rich source of MEVs, it is in our interest to investigate the biological potential of bovine MEVs. We have developed a protocol to obtain a pure MEV fraction from raw, untreated milk, validated, e.g. by the presence of well-described EV markers and absence of major milk contaminants such as casein and milk fat globules. In order to obtain more knowledge about the bioavailability of MEVs, we have investigated factors affecting in vitro uptake of MEVs in intestinal epithelium. Additionally, MEVs from processed milk have been isolated and compared to MEVs from unprocessed milk. Methods: MEVs from bovine milk were gently purified by size exclusion chromatography after an initial centrifugation step to remove milk fat and milk cells. For in vitro cell studies, isolated MEVs were specifically labelled with lactadherin marked with a fluorophore. Cellular uptake of MEVs was evaluated quantitatively by measuring total fluorescence on lysed cells. Results: Bovine MEVs were successfully labelled with fluorescent lactadherin. Quantitative measurements of cellular uptake of MEVs after incubation confirmed that MEVs are definitely internalized. Moreover, the investigations revealed that this uptake is time and temperature dependent. Several interventions were tested and evaluated in regard to cellular uptake. These include MEV concentration, temperature, simulated intestinal digestion conditions and the employment of different intestinal epithelial cell lines. Summary/Conclusion: A specific and non-invasive fluorescent labelling method was proven suitable to investigate bovine MEV uptake by different intestinal epithelial cell lines. In vitro cellular internalization of MEVs was confirmed, which indicates that MEVs definitely have the potential to convey bioactivity.
ISSN:2001-3078