Comparative study of commercial protocols for high recovery of high-purity mesenchymal stem cell-derived extracellular vesicle isolation and their efficient labeling with fluorescent dyes

Noriyasu Kamei; Haruka Nishimura; Atsushi Matsumoto; Riho Asano; Kanae Muranaka; Mahiro Fujita; Miina Takeda; Hiro Hashimoto; Mariko Takeda-Morishita

doi:10.1016/j.nano.2021.102396

Comparative study of commercial protocols for high recovery of high-purity mesenchymal stem cell-derived extracellular vesicle isolation and their efficient labeling with fluorescent dyes

Nanomedicine. 2021 Jul:35:102396. doi: 10.1016/j.nano.2021.102396. Epub 2021 Apr 15.

Authors

Noriyasu Kamei¹, Haruka Nishimura², Atsushi Matsumoto², Riho Asano², Kanae Muranaka², Mahiro Fujita², Miina Takeda², Hiro Hashimoto², Mariko Takeda-Morishita²

Affiliations

¹ Laboratory of Drug Delivery Systems, Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Kobe, Hyogo, Japan. Electronic address: noriyasu@pharm.kobegakuin.ac.jp.
² Laboratory of Drug Delivery Systems, Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Kobe, Hyogo, Japan.

PMID: 33864911
DOI: 10.1016/j.nano.2021.102396

Abstract

The extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) can be used as carriers for therapeutic molecules and drugs to target disordered tissues. This aimed to compare the protocols used for isolation of MSC-derived EVs by comparing EV collection conditions and three commercial purification kits. We also determined appropriate fluorescent dyes for labeling EVs. MSC-derived EVs were efficiently secreted during cell growth and highly purified by the phosphatidyl serine-based affinity kit. Although the EV membrane was more efficiently labeled with the fluorescent dye PKH67 compared to other probes, the efficiency was not enough to accurately analyze the endothelial cellular uptake of EVs. Results verified the easy protocol for isolating and fluorescently labeling EVs with commercial reagents and kits, but meanwhile, further modification of the protocol is required in order to scale up the amount of EVs derived from MSCs using fluorescent probes.

Keywords: Drug delivery; Exosome; Extracellular vesicle; Mesenchymal stem cell; Regenerative medicine.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Drug Carriers / chemistry*
Extracellular Vesicles* / chemistry
Extracellular Vesicles* / metabolism
Fluorescent Dyes / chemistry*
HeLa Cells
Humans
Mesenchymal Stem Cells* / chemistry
Mesenchymal Stem Cells* / metabolism
Staining and Labeling*

Substances

Drug Carriers
Fluorescent Dyes