Isolation of bovine milk-derived microvesicles carrying mRNAs and microRNAs

Hata, Taketoshi; Murakami, Kosuke; Nakatani, Hajime; Yamamoto, Yasunari; Matsuda, Tsukasa; Aoki, Naohito

doi:10.1016/j.bbrc.2010.04.135

Cited by 267 publications

(307 citation statements)

References 26 publications

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“…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]. This is probably due to the differences in the isolation method, such as acid treatment and ultracentrifugation processes.…”

Section: Discussionmentioning

confidence: 72%

“…According to the literature, mEVs have a biological function, such as an immunomodulatory effect [13]. In addition, mEVs and their cargos can be absorbed upon oral administration in humans [16].…”

Section: Introductionmentioning

confidence: 99%

“…To date, various isolation methods of mEVs have been developed, including ultracentrifugation [11,13,19] and size-exclusion chromatography [20]. However, none of them is suitable for the scalable production of mEVs.…”

Section: Introductionmentioning

confidence: 99%

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Biocompatibility of highly purified bovine milk‐derived extracellular vesicles

Somiya

Yoshioka²,

Ochiya³

2018

J of Extracellular Vesicle

182

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: Discussionmentioning

confidence: 72%

Section: Introductionmentioning

confidence: 99%

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Biocompatibility of highly purified bovine milk‐derived extracellular vesicles

Somiya

Yoshioka²,

Ochiya³

2018

J of Extracellular Vesicle

182

168

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“…Indeed, very recently Kosaka et al reported that breast milk contains miRNAs and these miRNAs may have potential biological functions in modulating cell growth and diffentiation [33]. Hata et al also showed that a considerable amount of mammary gland and immune-related miRNAs were present in the milk-derived microvesicles [34]. By comparing the miRNA profiling results obtained in cow milk and breast milk, we found that the majority of miRNAs, especially the immunespecific ones, are quite similar in cow milk and breast milk.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of microRNAs in raw milk during different periods of lactation, commercial fluid, and powdered milk products

et al. 2010

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Recent baby formula milk powder contamination incidents have shown that the classic markers or standards in milk quality control are insufficient in identifying "manipulated" poor-quality milk. In the present study, we demonstrated for the first time that cow milk contains large amounts of microRNAs (miRNAs) and that the unique expression profile of milk-specific miRNAs can serve as a novel indicator and possible new standard for the quality control of raw milk and milk-related commercial products, such as fluid milk and powdered formula milk. First, using Solexa sequencing, we systematically screened miRNA expression in raw milk and identified a total of 245 miRNAs in raw milk. Unlike other classic biomarkers whose expression levels are nearly identical at different periods of lactation, individual miRNAs can be significantly altered during lactation process, implicating that miRNAs may be a more accurate indicator to reflect the quality alteration of milk. Second, using TaqMan probe-based miRNA quantitative RT-PCR, we further identified seven miRNAs that have a relatively consistent expression throughout the lactation process, and more importantly, the expression profile of these seven milk-specific miRNAs can serve as an ideal biomarker for discriminating poor-quality or "manipulated" milk from pure raw milk, as well as for the quality control of commercial milk products, such as fluid milk and powdered formula milk. Together, our findings provide a basis for understanding the physiological role of milk miRNAs and a new potential standard for determining the quality of raw milk or milk-related commercial products.

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“…Knowledge about soluble MFG-E8 in milk is therefore limited. 18,27) In the present study, MFG-E8 secreted from insulin-stimulated HC11 cells was characterized with analyses by density-gradient ultracentrifugation and size-exclusion chromatography (Fig. 4A and B).…”

Section: Heterogeneity In the Mfg-e8 Distribution On Fat Globules In mentioning

confidence: 85%