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Technologies to Purify Exosomes from Bovine Milk and Alteration of Milk Exosomes Bioavailability in Humans and Rodents
Abstract
Bovine milk exosomes (BME) are bioavailable and transfer regulatory cargos across species boundaries. Thus, attracting considerable attention as bioactive compound and a vehicle in drug delivery. Exosome surface is highly glycosylated and has an impact on their biological properties and functions. However, role of glycosylation on surface of milk exosomes is elusive. The first chapter discusses the role of surface glycoproteins on milk exosomes in facilitating the exosome uptake and bioavailability in human intestinal cells and mice. Our findings suggest that modulation of surface glycans/glycoproteins suppressed the bioavailability and distribution of BME and their cargos post oral gavage. We also noted that based on mode of delivery and recipient cell, the bioavailability of surface glycan altered BME enhanced. In conclusion, our study contributes to a growing body of findings that bridge exosome glycosylation and functional studies. The second chapter expands upon the characterization of Exosome and RNA-depleted (ERD) and Exosome and RNA-sufficient (ERS) rodent diet based on BME, their cargos and bioavailability. We assessed the effect of milk ultrasonication on BME and their cargos. The results showed 85% lower bioavailable milk exosomes in the ERD diet than in the ERS diet. The uptake of exosomes isolated from ultrasonicated milk was 40% less efficient compared to exosomes isolated from non-ultrasonicated milk, in vitro and in vivo. Our work lays a foundation for conducting BME and RNA feeding studies in mice and humans. The third chapter explored the role of tangential flow filtration (TFF) to purify BME from cheesemaking waste. The large-scale protocols for the enrichment of BME from byproducts in dairy waste would be beneficial for utility of BME as an animal feed additive and drug delivery. We compared the size, morphology, protein and RNA cargos in EV purified by TFF to BME purified by serial ultracentrifugation. The result yielded heterogenous population of milk EVs. We conclude additional enrichment steps are needed to purify BMEs from cheese byproducts.
Subject Area
Nutrition
Recommended Citation
Sukreet, Sonal, "Technologies to Purify Exosomes from Bovine Milk and Alteration of Milk Exosomes Bioavailability in Humans and Rodents" (2021). ETD collection for University of Nebraska-Lincoln. AAI28258203.
https://digitalcommons.unl.edu/dissertations/AAI28258203