Isolation, identification, and characterization of novel nanovesicles

Abstract

Extracellular microvesicles (EVs) have been recognized for many potential clinical applications including biomarkers for disease diagnosis. In this study, we identified a major population of EVs by simply screening fluid samples with a nanosizer. Unlike other EVs, this extracellular nanovesicle (named HG-NV, HG-NV stands for HomoGenous nanovesicle as well as for Huang-Ge- nanovesicle) can be detected with a nanosizer with minimal in vitro manipulation and are much more homogenous in size (8–12 nm) than other EVs. A simple filtration platform is capable of separating HG-NVs from peripheral blood or cell culture supernatants. In comparison with corresponding exosome profiles, HG-NVs released from both mouse and human breast tumor cells are enriched with RNAs. Tumor derived HG-NVs are more potent in promoting tumor progression than exosomes. In summary, we identified a major subset of EVs as a previously unrecognized nanovesicle. Tumor cell derived HG-NVs promote tumor progression. Molecules predominantly present in breast tumor HG-NVs have been identified and characterized. This discovery may have implications in advancing both microvesicle biology research and clinical management including potential used as a biomarker.

Extraction and biomolecular analysis of dermal interstitial fluid collected with hollow microneedles

“…Using ImageJ Software 25 , exosome diameters purified from rat and human ISF were 140.4 ± 74.8 nm and 101.2 ± 43.9 nm, respectively, while exosomes purified from rat and human serum had diameters of 105.5 ± 42.0 nm and 103.2 ± 37.2 nm, respectively. Measured exosomes diameters agree with those established in the literature (30–150 nm) 23 – 27 . Using a nanoDrop spectrophotometer, we quantified the protein equivalent of purified exosome preparations from the ISF and serum of rats and humans.…”

“…Exosomes are recently described, cell-derived vesicles containing high concentrations of proteins and nucleic acid 22 , 23 . Exosomes appear to play important roles in intercellular signaling, and contain clinically relevant biomarkers for cancer and other illnesses 23 – 25 . We report here the first description to our knowledge of exosome content in dermal ISF, to our knowledge.…”

Boar seminal plasma exosomes maintain sperm function by infiltrating into the sperm membrane

“…To concentrate and identify boar seminal plasma exosomes, boar seminal plasma exosomes were isolated by ultracentrifugation using modified methods as previously described [ 10 , 14 ]. The exosome concentration of original seminal plasma (in Part I) were about 2×10 10 particles/mL as calculated by the NanoSight system.…”

“…Seminal plasma exosomes are membrane vesicles ranging from 30 to 120-nm diameter in size that are produced by organs in the male genital tracts including epididymis and prostate [ 10 , 14 , 15 ]. In boar, seminal plasma exosomes could inhibit the capacitation-dependent cholesterol efflux and mediate degradation of the 14-kD phosphorylated polypeptide of capacitated sperm [ 10 , 12 ].…”

Pathogenic roles of microvesicles in diabetic retinopathy

“…Microvesicles (MVs) are submicron membrane vesicles between 100 and 1000 nm that shed from the cell surface of both healthy and damaged cells [15] . By contrast, exosomes are smaller in diameter (between 40 and 100 nm) and are released during facultative or exocytosis [16] . The shedding of membrane MVs is a strictly regulated, cytoskeleton-dependent process that is enhanced by cytokines, the activation of apoptotic pathways, and reactive oxygen species [17] .…”