PHLIP – New phospholipid fractions in milk

Phospholipids (PL) are fundamental constituents of the membrane structures that defines the boundaries of all cells. In nutrition, special attention has been devoted to the PL sphingomyelin, which regulates cell growth, cell differentiation, and is important for development/function of the central nervous system. Sphingomyelin is not found in plants. Studies imply that fortification with sphingomyelin assist improved cognitive performance as well gut health and immunity early in life. Breast fed infants may have improved nutrition and development due to the presence of special PL and gangliosides in mothers’ milk.

By: Grith Mortensen

It is thought that the major part of the PL in cow’s milk originates from the milk fat globule membrane (MFGM). The research performed by the group at Aarhus University challenges this notion. Bovine milk contains ~1% PL, of which ~60% derive from MFGM. The rest of the PL is present in the skim milk and is suggested to be MFGM-remnants and is known as skim milk membranes. This nearly dogmatic perception is not valid anymore. Accordingly, another kind of PL structure appears to exist in milk, known as extracellular vesicles (EV). Milk EV holds an aqueous core, a high content of sfingomyelin and the molecular content differs considerable from MFGM. It is unclear to what extent current dairy process streams comprise EV and MFGM, which may lead to a heterogeneous PL-content and different biological activities.

This technological project is aiming to gain more information about milk lipid particles in dairy. The PHLIP-project has the following main objectives: 1) To clarify the occurrence of extracellular vesicles in milk fractions and reach a qualitative and quantitative description in relation to the appearance of MFG. 2) To establish a paradigm for analysis of EV and MFG in relevant dairy fractions.

Project period: January 2019-December 2021

Budget: 2.889.000 DKK

Financing: Milk Levy Fund, Aarhus University, Arla PhD fellowship

Project manager: Jan Trige Rasmussen

Institution: Department of Molecular Biology & Genetics, Aarhus University

Participants: Department of Molecular Biology & Genetics, Aarhus University and Arla Foods Ingredients

Publications

PhD progress report (Jan 2019). Maria Stenum Hansen. Biological Activities of Dairy Milk Extracellular Vesicles.

Thesis, Bachelor of Science (Jun 2019), Ida Schnack Eg Gadeberg, Assessment of biological effects of isolated bovine milk extracellular cells, 20 ECTS (eval. Jul 2019).

Thesis, Molecular Biological Project (June 2019), Susanne Rossen Ascanius, Investigation of extracellular vesicles’ biological effects on the intestinal immune system. 10 ECTS, Eval Jul 2019.

Oral presentation (3rd Oct. 2019), Susanne Rossen Ascanius, Maria Stenum Hansen, Jan Trige Rasmussen. Investigation of the biological effects of milk derived extracellular vesicles effect on cultured macrophages. 1st Annual symposium 2019, AU Hospital, Skejby.

Poster (Oct 2019), K. Blans, B. Petrat-Melin, X. Geng, L.B. Larsen, M.S. Ostenfeld, L. Wiking, J.T. Rasmussen. The composition of various phospholipid containing dairy fraction/powders is indicative for the origin of the comprising membrane material. 11th NIZO Dairy Conference, Papendal, NL.

Oral presentation (Nov 2019), Kristine Blans, Maria S. Hansen1, Marie S. Ostenfeld, and Jan T. Rasmussen. Isolation, characterization, and biological properties of milk extracellular vesicles. 16th International Symposium on Milk Genomics and Human Health – “Milk and health in a sustainability perspective”, Aarhus, 12-14 November 2019.

Poster (Nov 2019). Susanne Rossen Ascanius, Maria Stenum Hansen, Jan Trige Rasmussen. Investigation of the biological effects of milk derived extracellular vesicles effect on cultured macrophages. 16th International Symposium on Milk Genomics and Human Health – “Milk and health in a sustainability perspective”, Aarhus, 12-14 November 2019.

Thesis, Master of Science (Jun 2020), Susanne Rossen Ascanius, Investigation of biological effects of milk derived extracellular vesicles on macrophages. (Exam June 29, 2020).

Thesis, Molecular Biological Project (Jun 2020), Ida Schnack Eg Gadeberg, Immunological effect of extracellular vesicles in human macrophage-like cells,10 ECTS , Eval Jul 2020).

Thesis, Molecular Biological Project (Jun 2020), Pernille Moldrup Sørensen, Evaluation of protein markers specific for extracellular vesicles and milk fat globule membranes.10 ECTS. Eval Jul 2020.

Thesis, Bachelor of Science (Jun 2020), Caroline Vangsøe, Bovine milk derived extracellular vesicles influence on rotavirus infection, 20 ECTS (eval. Jul 2020).

Thesis, Bachelor of Science (Jun 2020), Rikke Halse Østergaard, Properties of bovine extracellular vesicles subjected to different treatments, 20 ECTS (eval. Jul 2020).

Maria S Hansen, Ida SE Gadegaard, Eva C Arnspang, Kristine Blans, Lene N Nejsum, Jan T Rasmussen. 2020. Specific and Non-Invasive Fluorescent Labelling of Extracellular Vesicles for Evaluation of Intracellular Processing by Intestinal Epithelial Cells. Biomedicines 8(7), 211.

Susanne R. Ascanius, Maria S. Hansen, Marie S. Ostenfeld, Jan T. Rasmussen (2021) Milk derived extracellular vesicles suppress inflammatory cytokine expression and nuclear factor-κB activation in lipopolysaccharide stimulated macrophages. Dairy, 2(2), 165-178: HTTPS://DOI.ORG/10.3390/DAIRY2020015