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A Method for the Isolation of Exosomes from Human and Bovine Milk

SCOPE: Milk provides a natural means of nutrient supply to infants. Exosomes are an important component of milk that are not only being studied for their promise in translational medicine but also in infant nutrition. They also play important roles in intercellular communication and immune function...

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Autores principales: Vaswani, Kanchan, Mitchell, Murray D., Holland, Olivia J., Qin Koh, Yong, Hill, Rebecca J., Harb, Tracy, Davies, Peter S. W., Peiris, Hassendrini
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6914892/
https://www.ncbi.nlm.nih.gov/pubmed/31885909
http://dx.doi.org/10.1155/2019/5764740
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author Vaswani, Kanchan
Mitchell, Murray D.
Holland, Olivia J.
Qin Koh, Yong
Hill, Rebecca J.
Harb, Tracy
Davies, Peter S. W.
Peiris, Hassendrini
author_facet Vaswani, Kanchan
Mitchell, Murray D.
Holland, Olivia J.
Qin Koh, Yong
Hill, Rebecca J.
Harb, Tracy
Davies, Peter S. W.
Peiris, Hassendrini
author_sort Vaswani, Kanchan
collection PubMed
description SCOPE: Milk provides a natural means of nutrient supply to infants. Exosomes are an important component of milk that are not only being studied for their promise in translational medicine but also in infant nutrition. They also play important roles in intercellular communication and immune function in mammary glands and are able to transfer their materials to the recipient. Therefore, the isolation of high-quality exosomes is an important aspect of exosome research. METHODS AND RESULTS: This study is a technical study, which provides a detailed methodology for the isolation and enrichment of exosomes from milk. In this study, we evaluate the suitability of using the exosome enrichment method that we have recently published for bovine milk, on human milk. We initially isolated extracellular vesicles from human and bovine milk on a fresh set of samples, using ultracentrifugation, and then exosomes were subsequently enriched via size exclusion chromatography (SEC). Following isolation and enrichment, exosomes from both species were characterized by particle concentration (nanoparticle tracking analysis, NTA), morphology (transmission electron microscopy, TEM), and the presence of exosomal markers (immunoblotting and mass spectrometry using information dependant acquisition (IDA)). The key exosomal characteristics of spherical/donut-shaped morphology, the presence of exosomal markers, e.g., FLOT-1 and the tetraspanins, CD9 and CD81), and particle concentration were confirmed in both human and bovine milk exosomes. CONCLUSION: We conclude that our robust exosome enrichment method, previously published for bovine milk, is suitable for use on human milk.
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spelling pubmed-69148922019-12-29 A Method for the Isolation of Exosomes from Human and Bovine Milk Vaswani, Kanchan Mitchell, Murray D. Holland, Olivia J. Qin Koh, Yong Hill, Rebecca J. Harb, Tracy Davies, Peter S. W. Peiris, Hassendrini J Nutr Metab Research Article SCOPE: Milk provides a natural means of nutrient supply to infants. Exosomes are an important component of milk that are not only being studied for their promise in translational medicine but also in infant nutrition. They also play important roles in intercellular communication and immune function in mammary glands and are able to transfer their materials to the recipient. Therefore, the isolation of high-quality exosomes is an important aspect of exosome research. METHODS AND RESULTS: This study is a technical study, which provides a detailed methodology for the isolation and enrichment of exosomes from milk. In this study, we evaluate the suitability of using the exosome enrichment method that we have recently published for bovine milk, on human milk. We initially isolated extracellular vesicles from human and bovine milk on a fresh set of samples, using ultracentrifugation, and then exosomes were subsequently enriched via size exclusion chromatography (SEC). Following isolation and enrichment, exosomes from both species were characterized by particle concentration (nanoparticle tracking analysis, NTA), morphology (transmission electron microscopy, TEM), and the presence of exosomal markers (immunoblotting and mass spectrometry using information dependant acquisition (IDA)). The key exosomal characteristics of spherical/donut-shaped morphology, the presence of exosomal markers, e.g., FLOT-1 and the tetraspanins, CD9 and CD81), and particle concentration were confirmed in both human and bovine milk exosomes. CONCLUSION: We conclude that our robust exosome enrichment method, previously published for bovine milk, is suitable for use on human milk. Hindawi 2019-12-03 /pmc/articles/PMC6914892/ /pubmed/31885909 http://dx.doi.org/10.1155/2019/5764740 Text en Copyright © 2019 Kanchan Vaswani et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Vaswani, Kanchan
Mitchell, Murray D.
Holland, Olivia J.
Qin Koh, Yong
Hill, Rebecca J.
Harb, Tracy
Davies, Peter S. W.
Peiris, Hassendrini
A Method for the Isolation of Exosomes from Human and Bovine Milk
title A Method for the Isolation of Exosomes from Human and Bovine Milk
title_full A Method for the Isolation of Exosomes from Human and Bovine Milk
title_fullStr A Method for the Isolation of Exosomes from Human and Bovine Milk
title_full_unstemmed A Method for the Isolation of Exosomes from Human and Bovine Milk
title_short A Method for the Isolation of Exosomes from Human and Bovine Milk
title_sort method for the isolation of exosomes from human and bovine milk
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6914892/
https://www.ncbi.nlm.nih.gov/pubmed/31885909
http://dx.doi.org/10.1155/2019/5764740
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