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In Journal of extracellular vesicles

The in vivo detection 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 vivo combined 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 cells in 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 EVs in vivo.

Kranich Jan, Chlis Nikolaos-Kosmas, Rausch Lisa, Latha Ashretha, Schifferer Martina, Kurz Tilman, Foltyn-Arfa Kia Agnieszka, Simons Mikael, Theis Fabian J, Brocker Thomas

2020-Jul-16

Extracellular Vesicles, apoptosis, dendritic cells, exosomes, irradiation, viral Infection