Journal
JOURNAL OF EXTRACELLULAR VESICLES
Volume 9, Issue 1, Pages -Publisher
WILEY
DOI: 10.1080/20013078.2020.1792683
Keywords
Extracellular Vesicles; exosomes; dendritic cells; viral Infection; irradiation; apoptosis
Categories
Funding
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) fellowship through the Graduate School of Quantitative Biosciences Munich (QBM)
- School of Life Sciences Weihenstephan, Technical University of Munich, Germany
- Graduate School QBM
- DFG within the Collaborative Research Centre (CRC) [1243]
- Helmholtz Association [ZT-I-0007]
- BMBF [01IS18036A, 01IS18053A]
- Chan Zuckerberg Initiative DAF (advised fund of Silicon Valley Community Foundation) [182835]
- DFG CRC [1054]
- DFG under Germany's Excellence Strategy within the framework of the Munich Cluster for Systems Neurology (EXC 2145 SyNergy) [390857198]
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Thein vivodetection of dead cells remains a major challenge due to technical hurdles. Here, we present a novel method, where injection of fluorescent milk fat globule-EGF factor 8 protein (MFG-E8)in vivocombined with imaging flow cytometry and deep learning allows the identification of dead cells based on their surface exposure of phosphatidylserine (PS) and other image parameters. A convolutional autoencoder (CAE) was trained on defined pictures and successfully used to identify apoptotic cellsin vivo. However, unexpectedly, these analyses also revealed that the great majority of PS(+)cells were not apoptotic, but rather live cells associated with PS(+)extracellular vesicles (EVs). During acute viral infection apoptotic cells increased slightly, while up to 30% of lymphocytes were decorated with PS(+)EVs of antigen-presenting cell (APC) exosomal origin. The combination of recombinant fluorescent MFG-E8 and the CAE-method will greatly facilitate analyses of cell death and EVsin vivo.
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