Exploring the adhesion properties of extracellular vesicles for functional assays.

Autor:	Pachane BC; Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.; Biochemistry and Molecular Biology Laboratory, Department of Physiological Sciences, Universidade Federal de São Carlos - UFSCar, São Carlos, SP, Brazil., Carlson B; Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Queen SE; Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA., Selistre-de-Araujo HS; Biochemistry and Molecular Biology Laboratory, Department of Physiological Sciences, Universidade Federal de São Carlos - UFSCar, São Carlos, SP, Brazil., Witwer KW; Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Oct 30. Date of Electronic Publication: 2024 Oct 30.
DOI:	10.1101/2024.10.28.620659
Abstrakt:	The "stickiness" of extracellular vesicles (EVs) can pose challenges for EV processing and storage, but adhesive properties may also be exploited to immobilize EVs directly on surfaces for various measurement techniques, including super-resolution microscopy. Direct adhesion to surfaces may allow examination of broader populations of EVs than molecular affinity approaches, which can also involve specialized, expensive affinity reagents. Here, we report on the interaction of EVs with borosilicate glass and quartz coverslips and on the effects of pre-coating coverslips with poly-L-lysine (PLL), a reagent commonly used to facilitate interactions between negatively charged surfaces of cells and amorphous surfaces. Additionally, we compared two mounting media conditions for super-resolution microscopy (SRM) imaging and used immobilized EVs for a B-cell interaction test. Our findings suggest that borosilicate glass coverslips immobilize EVs better than quartz glass coverslips. We also found that PLL is not strictly required for EV retention but contributes to the uniform distribution of EVs on borosilicate glass coverslips. Overall, these findings suggest that standard lab materials like borosilicate glass coverslips, with or without PLL, can be effectively used for the immobilization of EVs in specific imaging techniques. Competing Interests: Conflict of interest KWW is or has been an advisory board member of ShiftBio, Exopharm, NeuroDex, NovaDip, and ReNeuron; holds stock options with NeuroDex; and privately consults as Kenneth Witwer Consulting. Ionis Pharmaceuticals, Yuvan Research, and AgriSciX have sponsored or are sponsoring research in the Witwer laboratory.
Databáze:	MEDLINE
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