Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform

• Published in: International journal of molecular sciences Vol. 23; no. 15; p. 8544
• Main Authors: Breitwieser, Kai, Koch, Leon F., Tertel, Tobias, Proestler, Eva, Burgers, Luisa D., Lipps, Christoph, Adjaye, James, Fürst, Robert, Giebel, Bernd, Saul, Meike J.
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.08.2022; MDPI
• Subjects: Antibodies; CD63 antigen; CD81 antigen; CD9 antigen; Cell culture; characterization; Chemical compounds; Composition; Drug carriers; Drug delivery; Extracellular vesicles; Fibroblasts; Flow cytometry; Genotype & phenotype; Heterogeneity; Lymphocytes; Mesenchyme; Microscopy; Pharmacology; phenotype; Phenotypes; Populations; Proteins; small extracellular vesicles; Stem cells; Surface markers; tetraspanins; Vesicles
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23158544

Small extracellular vesicles (sEV) hold enormous potential as biomarkers, drug carriers, and therapeutic agents. However, due to previous limitations in the phenotypic characterization of sEV at the single vesicle level, knowledge of cell type-specific sEV signatures remains sparse. With the introduction of next-generation sEV analysis devices, such as the single-particle interferometric reflectance imaging sensor (SP-IRIS)-based ExoView R100 platform, single sEV analyses are now possible. While the tetraspanins CD9, CD63, and CD81 were generally considered pan-sEV markers, it became clear that sEV of different cell types contain several combinations and amounts of these proteins on their surfaces. To gain better insight into the complexity and heterogeneity of sEV, we used the ExoView R100 platform to analyze the CD9/CD63/CD81 phenotype of sEV released by different cell types at a single sEV level. We demonstrated that these surface markers are sufficient to distinguish cell-type-specific sEV phenotypes. Furthermore, we recognized that tetraspanin composition in some sEV populations does not follow a random pattern. Notably, the tetraspanin distribution of sEV derived from mesenchymal stem cells (MSCs) alters depending on cell culture conditions. Overall, our data provide an overview of the cell-specific characteristics of sEV populations, which will increase the understanding of sEV physiology and improve the development of new sEV-based therapeutic approaches.