Comparison of the Variability of Small Extracellular Vesicles Derived from Human Liver Cancer Tissues and Cultured from the Tissue Explants Based on a Simple Enrichment Method

Abstract

A potential use of small extracellular vesicles (sEVs) for diagnostic and therapeutic purposes has recently generated a great interest. sEVs, when purified directly from various tissues with proper procedures, can reflect the physiological and pathological state of the organism. However, the quality of sEV is affected by many factors during isolation, including separation of sEV from cell and tissues debris, the use of enzymes for tissue digestion, and the storage state of tissues. In the present study, we established an assay for the isolation and purification of liver cancer tissues-derived sEVs (tdsEVs) and cultured explants-derived sEVs (cedsEVs) by comparing the quality of sEVs derived from different concentration of digestion enzyme and incubation time. The nano-flow cytometry (NanoFCM) showed that the isolated tdsEVs by our method are purer than those obtained from differential ultracentrifugation. Our study thus establishes a simple and effective approach for isolation of high-quality sEVs that can be used for analysis of their constituents. Graphical abstract

Improving SERS biosensors for the analysis of ovarian cancer-derived small extracellular vesicles

“…2A1 and A2), which is consistent with the previously reported size range of the sEVs around 50–200 nm. 48 Besides, the concentrations of the sEVs from the OVCAR3 and A2780 cell lines were 1.5 × 10 8 particles per μL and 2.0 × 10 8 particles per μL, respectively (Fig. 2A1 and A2).…”

The Biological Effect of Small Extracellular Vesicles on Colorectal Cancer Metastasis

“…Existing techniques for the isolation of small extracellular vesicles, such as ultracentrifugation and size exclusion, fail to achieve ideal purity and yield. In addition, characterization methods and storage stability are also important issues in the clinical application of sEVs [ 85 , 86 ]. Further, due to the heterogeneity of sEVs, specific tools for differentiating sEVs from different intracellular origins are under development.…”

microRNAs profiling of small extracellular vesicles from midbrain tissue of Parkinson’s disease

“…For bdsEVs separation, we used a Tumor Dissociation Kit, human (Miltenyi Biotec, 130-095-929) following the manufacturer's protocol. The isolation was reported as described previously (Vella et al, 2017;Chen et al, 2021;Crescitelli et al, 2021). The midbrain tissues were cut into small pieces and weighed (~200 mg) with a cryostat, then placed in 2.2 mL DMEM/F-12 medium (Gibco, C11330500BT) containing 100 μL Enzyme H, 50 μL Enzyme R, and 12.5 μL Enzyme A, and incubated at 37°C for 15 min.…”