Bovine milk exosome proteome

Reinhardt, Timothy A.; Lippolis, John D.; Nonnecke, Brian J.; Sacco, Randy E.

doi:10.1016/j.jprot.2011.11.017

Cited by 216 publications

(238 citation statements)

References 46 publications

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“…The proteome analysis in this study (Table 2 and Supplementary Table 1) correlated well with a previous report showing that mEVs from bovine milk contain tetraspanins, heat shock proteins and other EV markers [28]. However, we could not detect some of the EV proteins, such as MFG-E8 (lactadherin), while this protein is frequently found in the mEV fraction [13,20].…”

Section: Discussionsupporting

confidence: 89%

Biocompatibility of highly purified bovine milk‐derived extracellular vesicles

Somiya

Yoshioka²,

Ochiya³

2018

J of Extracellular Vesicle

196

168

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Extracellular vesicles (EVs) deliver biologically active cargos from donor cells to recipient cells for intercellular communication. Since the existence of RNA cargo was discovered, EVs have been considered to be useful drug-delivery systems. Specifically, EVs from bovine milk (mEV) are one of the most promising platforms, since bovine milk is a scalable source of EVs for mass production. However, it is still difficult to isolate pure EVs from bovine milk owing to the complexity of raw materials. Furthermore, the biocompatibility and immunotoxicity of mEVs are still unclear. In this study, we developed a new method for isolating bovine milk-derived EVs by employing acid treatment and ultracentrifugation. Isolated mEVs are spherical in shape, measure 120 nm in diameter and contain typical EV marker proteins, such as tetraspanins. Compared with the previously reported method, our method can isolate purer mEVs. When mEVs are contacted with the mouse macrophage cell line Raw264.7, mEVs are readily taken up by the cells without a cytotoxic effect, suggesting that mEVs can deliver the cargo molecules into cells. While systemic administration of mEVs into mice resulted in the absence of systemic toxicity, certain types of cytokines were slightly induced. No anaphylaxis effect was observed after serial administration of mEVs in mice. Thus, mEVs isolated using our method are well tolerated in vivo and may be useful for the drug-delivery application.

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Section: Discussionsupporting

confidence: 89%

Biocompatibility of highly purified bovine milk‐derived extracellular vesicles

Somiya

Yoshioka²,

Ochiya³

2018

J of Extracellular Vesicle

196

168

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“…Milk is an important source of essential nutrients besides for lactating calves it also as a key raw material for human food preparations (Reinhardt et al 2012). All over the world people fulfill approximately 13% of their protein requirements from milk and dairy products.…”

Section: Introductionmentioning

confidence: 99%

Genotyping of Kappa-Casein Gene of Buffalo in Indonesian

Margawati

Volkandari²,

Indriawati³

et al. 2017

Proceedings of International Seminar on Livestock Production and Veterinary Technology

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Casein (KCN) is a milk protein with four variants of alpha S1, alpha S2, beta and kappa which has different allel of each. Kappa Casein (CSN3) gene lies at exon 4 associating with fat and protein milk contens. Genetic variation of CSN3 gene influences quality and composition of milk. The aim of this research was to genotype the CSN3 gene of Bufalos from several places in Indonesia. Amount of 29 buffalo samples was used in this research, those consisted of 15 collected from Medan, 3 samples of Banyuwangi, 10 samples of Baluran and one sample of South-East Nusa Tenggara (NTT). A Polymerase Chain Reaction-Restiction Fragment Length Polymorphism (PCR-RFLP) method was used for CSN3 gene analysis. The CSN3 gene was amplified at 56°C of annealing with 30 cycles and resulted a right fragment of 379 bp. The fragments were digested with HindIII at 37°C during for four hours for genotyping analysis. The result showed that BB genotypes were found at the locus of CSN3 gene in all samples. Those BB genotypes merged the pattern of digested fragments with sizes of 255 and 154 bp. This finding suggests that genotyping using HindIII reveals monomorphics of BB types accociated with milk content of casein.

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“…We determined that bovine milk exosomes contain mRNAs of major milk proteins and immune-related microRNAs [8]. More recently, Reinhardt et al [12] identified 2107 proteins in bovine milk exosomes. It is conceivable that bovine milk exosomes also play important roles, such as immune system maturation and responsiveness, in the growth of infants.…”

mentioning

confidence: 96%

Comparison of Methods for Isolating Exosomes from Bovine Milk

et al. 2012

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ABSTRACT. Four methods were evaluated for isolating exosomes from bovine milk: (1) ExoQuick precipitation, (2) ultracentrifugation with ExoQuick precipitation, (3) ultracentrifugation with density gradient centrifugation, and (4) human milk exosome isolation. Methods 1 and 4 failed due to differences between bovine and human milk. Exosomes were efficiently isolated by ultracentrifugation with either ExoQuick precipitation (method 2) or density gradient centrifugation (method 3). The highest yield of exosomes was achieved using ultracentrifugation with ExoQuick precipitation, whereas higher quality exosome isolation with intact morphological structures was achieved by ultracentrifugation with density gradient centrifugation.

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Bovine milk exosome proteome

Cited by 216 publications

References 46 publications

Biocompatibility of highly purified bovine milk‐derived extracellular vesicles

Biocompatibility of highly purified bovine milk‐derived extracellular vesicles

Genotyping of Kappa-Casein Gene of Buffalo in Indonesian

Comparison of Methods for Isolating Exosomes from Bovine Milk

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