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Increased Yield of Extracellular Vesicles after Cytochalasin B Treatment and Vortexing.

ABSTRACT: Extracellular vesicles (EVs) are promising therapeutic instruments and vectors for therapeutics delivery. In order to increase the yield of EVs, a method of inducing EVs release using cytochalasin B is being actively developed. In this work, we compared the yield of naturally occurring extracellular vesicles and cytochalasin B-induced membrane vesicles (CIMVs) from mesenchymal stem cells (MSCs). In order to maintain accuracy in the comparative analysis, the same culture was used for the isolation of EVs and CIMVs: conditioned medium was used for EVs isolation and cells were harvested for CIMVs production. The pellets obtained after centrifugation 2300× g, 10,000× g and 100,000× g were analyzed using scanning electron microscopy analysis (SEM), flow cytometry, the bicinchoninic acid assay, dynamic light scattering (DLS), and nanoparticle tracking analysis (NTA). We found that the use of cytochalasin B treatment and vortexing resulted in the production of a more homogeneous population of membrane vesicles with a median diameter greater than that of EVs. We found that EVs-like particles remained in the FBS, despite overnight ultracentrifugation, which introduced a significant inaccuracy in the calculation of the EVs yield. Therefore, we cultivated cells in a serum-free medium for the subsequent isolation of EVs. We observed that the number of CIMVs significantly exceeded the number of EVs after each step of centrifugation (2300× g, 10,000× g and 100,000× g) by up to 5, 9, and 20 times, respectively.

SUBMITTER: Kurbangaleeva SV

PROVIDER: S-EPMC10047179 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Increased Yield of Extracellular Vesicles after Cytochalasin B Treatment and Vortexing.

Kurbangaleeva Sirina V SV Syromiatnikova Valeriia Y VY Prokopeva Angelina E AE Rogov Aleksey M AM Khannanov Artur A AA Rizvanov Albert A AA Gomzikova Marina O MO

Current issues in molecular biology 20230315 3

Extracellular vesicles (EVs) are promising therapeutic instruments and vectors for therapeutics delivery. In order to increase the yield of EVs, a method of inducing EVs release using cytochalasin B is being actively developed. In this work, we compared the yield of naturally occurring extracellular vesicles and cytochalasin B-induced membrane vesicles (CIMVs) from mesenchymal stem cells (MSCs). In order to maintain accuracy in the comparative analysis, the same culture was used for the isolatio ...[more]

PMID: 36975528

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Increased Yield of Extracellular Vesicles after Cytochalasin B Treatment and Vortexing.

Publications

Increased Yield of Extracellular Vesicles after Cytochalasin B Treatment and Vortexing.

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