Characterization of extracellular vesicles derived from cardiac cells in an in vitro model of preconditioning

Journal of Extracellular Vesicles
Sebastian BoroschSandra Kraemer

Abstract

Preconditioning is a promising technique to protect the heart from ischaemia-reperfusion injury. In this context, the crosstalk between different cardiac cell types and especially the exchange of cardioprotective mediators has come into the focus of current research. Recently, extracellular vesicles (EVs), nano-sized structures, emerged as possible communication mediators. They are taken up by recipient cells and can alter gene expression or activate intracellular signal cascades. It has been shown that all cardiac cell types are able to secrete EVs, but so far the influence of an in vitro preconditioning stimulus on EV concentration and composition has not been investigated. Therefore, we stimulated primary cardiac myocytes and fibroblasts from neonatal rats, as well as H9c2 cells, with two known in vitro preconditioning stimuli: hypoxia or isoflurane. EVs were isolated from cell culture supernatants 48 h after stimulation by differential centrifugation and size exclusion chromatography. They were characterized by transmission electron microscopy, tunable resistive pulse sensing, miRNA array and Western blot analysis. The detected EVs had the typical cup-shaped morphology and a size of about 150 nm. No significant differences ...Continue Reading

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Citations

Dec 21, 2018·Journal of Applied Physiology·Glenn D WadleyBianca C Bernardo
Nov 2, 2019·The European Respiratory Journal·Abdelnaby KhalyfaBharati Prasad
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Methods Mentioned

BETA
size exclusion chromatography
electron microscopy
transmission electron microscopy
chips

Software Mentioned

ImageJ
GraphPad Prism
GraphPad
MRI _ Wound _ Healing _ Tool
AltAnalyze
Izon Control Suite
expression console
Image Lab

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