Stability of erythrocyte-derived nanovesicles assessed by light scattering and electron microscopy

Božič, Darja (avtor), Hočevar, Matej (avtor), Kisovec, Matic (avtor), Pajnič, Manca (avtor), Pađen, Ljubiša (avtor), Jeran, Marko (avtor), Bedina Zavec, Apolonija (avtor), Podobnik, Marjetka (avtor), Kogej, Ksenija (avtor), Iglič, Aleš (avtor), Kralj-Iglič, Veronika (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: 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
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: ZF - Zdravstvena fakulteta ; FE - Fakulteta za elektrotehniko; FKKT - Fakulteta za kemijo in kemijsko tehnologijo ; MF - Medicinska fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2021
• Št. strani: 28 str.
• Številčenje: Vol. 22, iss. 23, art. 12772
• PID: 20.500.12556/RUL-133687
• UDK: 577
• ISSN pri članku: 1422-0067
• DOI: 10.3390/ijms222312772
• COBISS.SI-ID: 88130563
• Datum objave v RUL: 09.12.2021

Gradivo je del revije

• Naslov: International journal of molecular sciences
• Skrajšan naslov: Int. j. mol. sci.
• Založnik: MDPI
• ISSN: 1422-0067
• COBISS.SI-ID: 2779162

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.
• Začetek licenciranja: 01.12.2021

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: sipanje svetlobe, karakterizacija veziklov, stabilnost veziklov, referenčni material zunajceličnih veziklov, nanovezikli, celični vezikli, oblika veziklov, vrstična elektronska mikroskopija zunajceličnih veziklov, krioelektronska mikroskopija zunajceličnih veziklov