Extracellular Vesicles of Mesenchymal Stem Cells Are More Effectively Accessed through Polyethylene Glycol-Based Precipitation than by Ultracentrifugation

Lei Jia; Bo Li; Cong Fang; Xiaoyan Liang; Yingjun Xie; Xiaofang Sun; Wen Wang; Lei Zheng; Ding Wang

doi:10.1155/2022/3577015

Extracellular Vesicles of Mesenchymal Stem Cells Are More Effectively Accessed through Polyethylene Glycol-Based Precipitation than by Ultracentrifugation

Stem Cells Int. 2022 Sep 6:2022:3577015. doi: 10.1155/2022/3577015. eCollection 2022.

Authors

Lei Jia^{1

2

3}, Bo Li^{1

2}, Cong Fang³, Xiaoyan Liang³, Yingjun Xie^{4

5}, Xiaofang Sun^{4

5}, Wen Wang^{1

2

6}, Lei Zheng^{1

2}, Ding Wang^{4

5}

Affiliations

¹ The First School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China.
² Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
³ Reproductive Medicine Center, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou 510275, China.
⁴ Department of Obstetrics and Gynecology, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150 Guangdong, China.
⁵ Guangdong Provincial Key Laboratory of Major Obstetric Diseases, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150 Guangdong, China.
⁶ School of Engineering and Materials Science, Queen Mary University of London, London E1 4NS, UK.

Abstract

Extracellular vesicles (EVs) have been identified as cell-cell communication agents, and EVs derived from mesenchymal stem cells (MSCs) exhibit therapeutic effects similar to those of the cells of origin. Precipitation methods have been used extensively for EV harvests, such as UC- (ultracentrifugation-) or PEG- (polyethylene glycol-) based methods, and the difference in EVs derived from MSCs by UC and PEG is not fully understood. We harvested EVs from amniotic fluid MSCs (AF-MSCs) by UC- or PEG-based precipitation methods and conducted a comparison study of those EVs derived by the two methods: output, RNA, and protein expression of EVs and EV biological reaction in a THP-1-cell model of LPS induction, which was considered an infection model. There was no difference in morphology, size, or specific marker-positive ratio of PEG-EVs and UC-EVs, but PEG obtained more EV particles, protein, and RNA than the UC method. In our THP-1 model of LPS induction, MSC-EVs did not lead to a change in protein expression but inhibited the LPS-induced increase in cytokine secretion. UC-EVs were more effective for TNF-α inhibition, and PEG-EVs were more effective for IL10 inhibition. Thus, our findings provide evidence that PEG-based precipitation is a more efficient mesenchymal stem cell-extracellular vesicle-derived method than UC.