文献类型：期刊文献

英文题名：Optimizing Conditions of Polyethylene Glycol Precipitation for Exosomes Isolation From MSCs Culture Media for Regenerative Treatment

作者：Yu, Junjun[1];Huang, Daqiang[1];Liu, Hanwen[1];Cai, Haibo[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China

年份：2024

卷号：19

期号：9

外文期刊名：BIOTECHNOLOGY JOURNAL

收录：;EI(收录号：20243917092769);WOS:【SCI-EXPANDED(收录号:WOS:001315056800001)】；

语种：英文

外文关键词：exosomes; miRNAs; polyethylene glycol; proteins

摘要：Mesenchymal stem cell (MSC)-derived exosomes, as a cell-free alternative to MSCs, offer enhanced safety and significant potential in regenerative medicine. However, isolating these exosomes poses a challenge, complicating their broader application. Commonly used methods like ultracentrifugation (UC) and tangential flow filtration are often impractical due to the requirement for costly instruments and ultrafiltration membranes. Additionally, the high cost of commercial kits limits their use in processing large sample volumes. Polyethylene glycol (PEG) precipitation offers a more convenient and cost-effective alternative, but there is a critical need for optimized and standardized isolation protocols using PEG precipitation across different cell types and fluids to ensure consistent quality and yield. In this work, we optimized the PEG precipitation method for exosomes isolation and compared its effectiveness to two commonly used methods: UC and commercial exosome isolation kits (ExoQuick). The recovery rate of the optimized PEG method (about 61.74%) was comparable to that of the commercial ExoQuick kit (about 62.19%), which was significantly higher than UC (about 45.80%). Exosome cargo analysis validated no significant differences in miRNA and protein profiles associated with the proliferation and migration of exosomes isolated by UC and PEG precipitation, which was confirmed by gap closure and CCK8 assays. These findings suggest that PEG-based exosomes isolation could be a highly efficient and high-quality method and may facilitate the development of exosome-based therapies for regenerative medicine. The conditions for the separation of exosomes by polyethylene glycol (PEG) polymerization precipitation were optimized, and the recovery rate was comparable to the commercial kit ExoQuick and higher than that of the traditional ultracentrifugation (UC) method. Further analyzing the contents of exosomes, the miRNAs and proteins associated with proliferation and migration in exosomes isolated by UC and PEG methods were similar, which was confirmed by cell function tests in vitro. image