Fig. 1: Schematic representation of the generation of nano plasma membrane vesicles (nPMVs), their characterization, and in vitro/in vivo studies for comparison to native extracellular vesicles (EVs). | Communications Biology

Fig. 1: Schematic representation of the generation of nano plasma membrane vesicles (nPMVs), their characterization, and in vitro/in vivo studies for comparison to native extracellular vesicles (EVs).

From: High efficiency preparation of monodisperse plasma membrane derived extracellular vesicles for therapeutic applications

Fig. 1: Schematic representation of the generation of nano plasma membrane vesicles (nPMVs), their characterization, and in vitro/in vivo studies for comparison to native extracellular vesicles (EVs).

Donor cells are treated with chemical stressors (i.e., paraformaldehyde (PFA), dithiothreitol (DTT)) to induce apoptosis, which results in membrane blebbing and the production of ca. 1–3 µm sized apoptotic bodies (ApoBDs), namely giant plasma membrane vesicles (GPMVs). GPMVs are isolated and processed into unilamellar nPMVs through extrusion and purification to obtain a defined, monodisperse, and reproducible size distribution. nPMVs possibly differ in cargo and protein composition. The production rate and yield of the nPMV and native EV preparation method were evaluated. nPMVs were thereafter directly compared to native EVs using physico-chemical methods, 2D- and 3D-cryo-TEM, proteomics, lipidomics, in vitro recipient cell interactions using several human cell lines (i.e., Huh7 cells and THP-1 M0 macrophages), and biodistribution in transgenic (Tg) zebrafish larvae (ZFL) (i.e., Tg(kdrl:EGFP), Tg(mpeg:Kaede) (short for Tg(mpeg1:Gal4:UAS:Kaede)) as an in vivo vertebrate model.

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