Pregled bibliografske jedinice broj: 1211861
Unveiling the Native Morphology of Extracellular Vesicles from Human Cerebrospinal Fluid by Atomic Force and Cryogenic Electron Microscopy
Unveiling the Native Morphology of Extracellular Vesicles from Human Cerebrospinal Fluid by Atomic Force and Cryogenic Electron Microscopy // Biomedicines, 10 (2022), 6; 1251, 10 doi:10.3390/biomedicines10061251 (međunarodna recenzija, članak, znanstveni)
CROSBI ID: 1211861 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Unveiling the Native Morphology of Extracellular
Vesicles from Human Cerebrospinal Fluid by Atomic
Force and Cryogenic Electron Microscopy
Autori
Kurtjak, Mario ; Kereïche, Sami ; Klepac, Damir ; Križan, Hrvoje ; Perčić, Marko ; Krušić Alić, Vedrana ; Lavrin, Teja ; Lenassi, Metka ; Wechtersbach, Karmen ; Kojc, Nika ; Vukomanović, Marija ; Zrna, Siniša ; Biberić, Maša ; Domitrović, Robert ; Grabušić, Kristina ; Malenica, Mladenka
Izvornik
Biomedicines (2227-9059) 10
(2022), 6;
1251, 10
Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni
Ključne riječi
extracellular vesicles ; cerebrospinal fluid ; size-exclusion chromatography ; atomic force microscopy ; cryogenic transmission electron microscopy ; morphology 1. Introduction
Sažetak
Extracellular vesicles (EVs) are membranous structures in biofluids with enormous diagnostic/prognostic potential for application in liquid biopsies. Any such downstream application requires a detailed characterization of EV concentration, size and morphology. This study aimed to observe the native morphology of EVs in human cerebrospinal fluid after traumatic brain injury. Therefore, they were separated by gravity- driven size-exclusion chromatography (SEC) and investigated by atomic force microscopy (AFM) in liquid and cryogenic transmission electron microscopy (cryo-TEM). The enrichment of EVs in early SEC fractions was confirmed by immunoblot for transmembrane proteins CD9 and CD81. These fractions were then pooled, and the concentration and particle size distribution were determined by Tunable Resistive Pulse Sensing (around 1010 particles/mL, mode 100 nm) and Nanoparticle Tracking Analysis (around 109 particles/mL, mode 150 nm). Liquid AFM and cryo-TEM investigations showed mode sizes of about 60 and 90 nm, respectively, and various morphology features. AFM revealed round, concave, multilobed EV structures ; and cryo-TEM identified single, double and multi- membrane EVs. By combining AFM for the surface morphology investigation and cryo-TEM for internal structure differentiation, EV morphological subpopulations in cerebrospinal fluid could be identified. These subpopulations should be further investigated because they could have different biological functions.
Izvorni jezik
Engleski
POVEZANOST RADA
Projekti:
--uniri-biomed-18-279 - Uspostava protokola na Sveučilištu u Rijeci za primjenu nanotehnoloških metoda: mikroskopije atomskih sila i skenirajućeg elektronskog mikroskopa prilikom vizualizacije i karakterizacije egzosoma (Malenica Staver, Mladenka; Malenica Staver, Mladenka) ( CroRIS)
HRZZ-IP-2019-04-1511 - Identifikacija cirkulirajućih biomarkera neurološkog oporavka u bolesnika s ozljedom mozga (CiMozak / CirBrain) (Grabušić, Kristina, HRZZ - 2019-04) ( CroRIS)
Profili:
Marko Perčić
(autor)
Mladenka Malenica
(autor)
Damir Klepac
(autor)
Kristina Grabušić
(autor)
Hrvoje Križan
(autor)
Robert Domitrović
(autor)
Citiraj ovu publikaciju:
Časopis indeksira:
- Current Contents Connect (CCC)
- Web of Science Core Collection (WoSCC)
- Science Citation Index Expanded (SCI-EXP)
- SCI-EXP, SSCI i/ili A&HCI
- Scopus