The network is a local initiative started at Aarhus University to bring together researchers and students working in the highly interdiscplinary field of extracellular vesicles (EVs). The aim is to boost our research at large by joining forces across different departments/centres through collaborations and student education.
Key words: Bioimaging, EVomics, EV Characterization, Tissue Regeneration, Macrophage & Endothelial Biology
miRNA
Universitetsbyen 81
We study EVs as the new frontier of cell-to-cell communication. For example, by which mechanisms can EVs find the target recipient cells via the bloodstream? What kind of messages are conveyed to regulate/support the recipient cells? With zebrafish as our little partners, we seek answers to these questions by nanoscience approaches, bioinformatics and 4D imaging of live transgenic embryos.
We have two approaches: fluorescent labelling of endogenous EVs and microinjection of (fluorescently labelled) EVs including, but not limited to, human-derived EVs. In both cases, EVs are imaged in live zebrafish embryos using optical microscopes to study the biodistribution, blood clearance kinetics, and interactions with cells of interest (e.g. macrophages and endothelial cells). Induction of TNF-alpha can also be visualized in real-time as an indicator of macrophage polarization in response to EVs. Collaborations on tissue injury models and EV injections are welcome.
ChromoTek’s GFP-trap Dynabeads for EV isolation from tissues (zebrafish embryos).
Through Embion. Cryo-electron microscopy (cryo-EM) and cryo-correlative light-electron microscopy (cryo-CLEM) for imaging EVs at ultrastructural resolution.
EV characterization
Høegh-Guldbergs Gade 10
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Capable of resolving particles down to 160 nm on scatter. Johann Mar Gudbergsson has experience in analysing EVs with this.
Capable of resolving particles down to 100 nm only by scatter but smaller when using fluorescence. Yuya Hayashi has experience in analysing EVs with this.
Capable of resolving particles down to 100 nm only by scatter but smaller when using fluorescence.
A cell sorter which has small particle detection mode enabling it to resolve/sort particles down to 100-160 nm – but the instrument is very new and we did not have time to test it yet. In theory, it should be possible to sort EVs on this.
miRNA
Gustav Wieds Vej 14
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Nanoimager fluorescent and super-resolution microscope. Can image in both epifluorescence, total internal reflection (TIRF) and HILO. The microscope has four laser lines 405 nm, 488 nm, 561 nm, and 640 nm. The microscope can image in real-time ideal for tracking of single molecules and is equipped with a temperature controller and a micro fluiding pumping system. Read more here (https://oni.bio/nanoimager/).
Contact person for ONI: Mette Malle <malle@inano.au.dk>
OBS: the microscope is not trivial to use and Mette is happy to collaborate and/or help with everything from guiding which fluorophores should be used, microscope surface preparation and imaging conditions.
Nanoparticle tracking analysis (NTA) for size and concentration determination (single-particle approach).
Dynamic light scattering (DLS) for determination of size distributions (ensemble approach, intensity biased).
miRNA, Protein
Palle Juul-Jensens Boulevard 45
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In vivo (pre-)labelling of EVs, engineered labelling of EVs, and engineered loading of therapeutic proteins into EVs
Augmented COlorimetric NANoplasmonic (CONAN) method for determining purity and concentration of EVs using gold nanoparticles.
RAW264.7, SIM-A9, THP-1, HEK293T, IPEC-J2, PBMCs, Monocytes, Adipose derived stem cells (ASCs)
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Palle Juul-Jensens Boulevard 45
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For routine EV isolation.
Nanoparticle tracking analysis (NTA) for size and concentration determination (single-particle approach). Available at Blood & Biochemistry, AUH.
miRNA
Gustav Wieds Vej 14
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Omiics can offer RNA sequencing service to EV samples and also offer suggestions on EV samples preparation for RNA sequencing. We have established bioinformatics pipeline for analyzing the small RNAs (miRNAs, tRNA derived small RNAs and other small RNAs) and long-RNAs (mRNAs, circular RNAs and long non-coding RNAs) in EV samples. The analysis include: quantification of different types of RNAs, differential expression analysis, pathway analysis and so on.
