Comparison of separation methods for tissue‐derived extracellular vesicles in the liver, heart, and skeletal muscle

Abstract

Extracellular vesicles (EVs), which are nanosized vesicles released by cells as intracellular messengers, have high potential as biomarkers. EVs are usually collected from in vitro sources, such as cell culture media or biofluids, and not from tissues. Techniques enabling direct collection of EVs from tissues will extend the applications of EVs. We compared methods for separating EVs from solid liver, heart, and skeletal muscle. Compared with a precipitation method, an ultracentrifugation‐based method for collection of EVs from solid tissues yielded a higher proportion of EVs positive for EV‐related markers, with minimum levels of intracellular organelle‐related markers. Some tissue‐specific modifications, such as a sucrose cushion step, may improve the yield and purity of the collected EVs.

Biogenic nanoparticles from liquid and solid matrices: biochemical and biophysical properties of Extracellular Vesicles-enriched samples from human plasma and skeletal muscle tissue.

“…Similarly, fractions 6 to 9 of Tissue 16k (Figure 3A) and Tissue 118k (Figure 3B) samples, after DSG processing, resulted enriched in EV markers and in SkM-EV markers. The SkM-EV marker Caveolin-3 was not detectable in plasma samples (Fig.2A,B), indicating that, with this experimental conditions, this protein is a specific SkM-EV marker 10,12,48 .…”

“…The SkM-EV marker Caveolin-3 was not detectable in plasma samples (Fig. 2A,B), indicating that, with this experimental conditions, this protein is a specific SkM-EV marker 10,12,48 .…”

Proteomic and miRNA Profiles of Exosomes Derived from Myometrial Tissue in Laboring Women

“…Exosomes isolated from skeletal muscle tissue have been reported [ 18 , 19 ]; however, research on the uterine myometrial exosomes is lacking. In this study, we successfully isolated exosomes from the human myometrium and characterized its proteins and miRNA cargoes through liquid chromatography–tandem mass spectrometry (LC-MS/MS) and miRNA sequencing.…”

Research advances of tissue-derived extracellular vesicles in cancers

“…UC samples contained more EVs of smaller size than precipitation. The calnexin, a protein associated with endoplasmic reticulum, was detected in Ti-EVs isolated by precipitation, but not in UC samples, indicating that Ti-EVs isolated by precipitation contained non-EV contamination (Matejovič et al 2021 ). UC is currently the most popular EVs isolation method (Gardiner et al 2016 ).…”