20.500.12556/RUL-133687 Stability of erythrocyte-derived nanovesicles assessed by light scattering and electron microscopy Extracellular vesicles (EVs) are gaining increasing amounts of attention due to their potential use in diagnostics and therapy, but the poor reproducibility of the studies that have been conducted on these structures hinders their breakthrough into routine practice. We believe that a better understanding of EVs stability and methods to control their integrity are the key to resolving this issue. In this work, erythrocyte EVs (hbEVs) were isolated by centrifugation from suspensions of human erythrocytes that had been aged in vitro. The isolate was characterised by scanning (SEM) and cryo-transmission electron microscopy (cryo-TEM), flow cytometry (FCM), dynamic/static light scattering (LS), protein electrophoresis, and UV-V spectrometry. The hbEVs were exposed to various conditions (pH (4–10), osmolarity (50–1000 mOsm/L), temperature (15–60 °C), and surfactant Triton X-100 (10–500 μM)). Their stability was evaluated by LS by considering the hydrodynamic radius (R$_h$), intensity of scattered light (I), and the shape parameter (ρ). The morphology of the hbEVs that had been stored in phosphate-buffered saline with citrate (PBS–citrate) at 4 °C remained consistent for more than 6 months. A change in the media properties (50–1000 mOsm/L, pH 4–10) had no significant effect on the R$_h$ (=100–130 nm). At pH values below 6 and above 8, at temperatures above 45 °C, and in the presence of Triton X-100, hbEVs degradation was indicated by a decrease in I of more than 20%. Due to the simple preparation, homogeneous morphology, and stability of hbEVs under a wide range of conditions, they are considered to be a suitable option for EV reference material. light scattering vesicle characterization vesicle stability extracellular vesicle reference material nanovesicles cellular vesicles vesicle shape scanning electron microscopy of extracellular vesicles cryo-electron microscopy of extracellular vesicles sipanje svetlobe karakterizacija veziklov stabilnost veziklov referenčni material zunajceličnih veziklov nanovezikli celični vezikli oblika veziklov vrstična elektronska mikroskopija zunajceličnih veziklov krioelektronska mikroskopija zunajceličnih veziklov true false true Angleški jezik Slovenski jezik Članek v reviji 2021-12-09 10:03:24 2021-12-09 10:03:25 2023-05-04 14:24:48 0000-00-00 00:00:00 2021 0 0 28 str. iss. 23, art. 12772 Vol. 22 2021 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 1422-0067 10.3390/ijms222312772 88130563 ijms-22-12772-v3.pdf ijms-22-12772-v3.pdf 1 A39E3A98370B883D574738574EA66AAD 53bf5990e468d8a5fd76436f6dba2db93fb2f8419a5a2754597e57a8d3c412b6 708118e5-d67a-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=156484 https://www.mdpi.com/1422-0067/22/23/12772 1 c2619459-58ce-11ec-abdb-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=151961 Zdravstvena fakulteta Fakulteta za elektrotehniko Fakulteta za kemijo in kemijsko tehnologijo Medicinska fakulteta 0 0 0