Comparison of qNANO results from the isolation of extracellular microvesicles with the theoretical model

Abstract

Objectives: Extracellular vesicles (EVs) are heterogeneous membrane vesicles in diameter of 30-5000 nm, that transport proteins, non-coding RNAs (miRNAs), lipids and metabolites. Major populations include exosomes, ectosomes and apoptotic bodies. The purpose of this study was to compare the distribution of EVs obtained under different conditions of differential centrifugation, including ultracentrifugation, with the results developed based on a theoretical model. Methods: Immortalized endothelial cell line that expresses h-TERT (human telomerase) was used to release of EVs: microvascular TIME. EVs were isolated from the culture medium at different centrifugation parameters. The size distribution of the EVs was measured using TRPS technology on a qNano instrument.

Revealing sphingolipids composition in extracellular vesicles and paternal β-cells after persistent hyperglycemia

“…As demonstrated by Mathilde Mathieu et al, both tetraspanins CD9 and CD63 can be released in small ectosomes formed at the plasma membrane, a finding that our results corroborate [63]. Similar observations were reported in studies performed on the immortalized endothelial cells [64].…”

“…In this study, the presence of both large and small EVs in isolated samples was confirmed not only through CryoTEM and TEM analyses but also via spectral flow cytometry, which is based on the detection of specific markers: CD9, CD63, CD81, and annexin-V. As demonstrated by Mathilde Mathieu et al, both tetraspanins CD9 and CD63 can be released in small ectosomes formed at the plasma membrane, a finding that our results corroborate [63]. Similar observations were reported in studies performed on the immortalized endothelial cells [16].…”

“…EVs can be divided into several subpopulations that differ in biogenesis, size, cargo and function [15]. Currently, the main EV populations recognize large EVs (150 – 1000 nm) and small EVs (50 – 150 nm) [16]. Their size is of great importance because it can be used to assume the biogenesis and function of EVs, while large EVs are directly secreted from the cell membrane, small EVs accumulate in the lumen of late endosomal compartments and are released into the extracellular space after endosome fusion with the cell membrane [17,18].…”

miRNA Signature of Urine Extracellular Vesicles Shows the Involvement of Inflammatory and Apoptotic Processes in Diabetic Chronic Kidney Disease

“…A large age variation within the group of diabetic patients (45–88) and a difference in average age between the control group (median = 50) and patients (median = 82.5) may also affect the physicochemical properties of EVs. In our previous study, we observed age has a stronger contribution in patients with CKD compared to patients without kidney disease [ 58 ]. In this study, the strong correlation of age with selected miRNAs was demonstrated.…”

“…We used the cellulose dialysis membrane with 1000 kDa MWCO to concentrate EVs from the urine sample, nevertheless the urinary EV sample may be slightly contaminated by Tamm Horsfall Protein (about 4% is retained), which has been visualized in our previous study using SEM imaging methods [ 59 ]. It has been described in previous articles that the applied urinary EV isolation method may also affect the yields of miRNAs [ 58 ]. After the low-pressure filtration procedure, the isolated uEV samples were frozen at -80°C.…”