Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography

Abstract As functional derivatives of mesenchymal stem cells (MSCs), small extracellular vesicles (sEVs) have garnered significant attention and application in regenerative medicine. However, the technical limitations for large‐scale isolation of sEVs and their heterogeneous nature have added comple...

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Main Authors: Wei Liu, Xinyu Wang, Yating Chen, Jiapei Yuan, Huiyu Zhang, Xin Jin, Yuying Jiang, Junjing Cao, Zibin Wang, Shuo Yang, Bingwei Wang, Tinghe Wu, Jing Li
Format: Article
Language:English
Published: Wiley 2025-01-01
Series:Journal of Extracellular Vesicles
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Online Access:https://doi.org/10.1002/jev2.70029
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author Wei Liu
Xinyu Wang
Yating Chen
Jiapei Yuan
Huiyu Zhang
Xin Jin
Yuying Jiang
Junjing Cao
Zibin Wang
Shuo Yang
Bingwei Wang
Tinghe Wu
Jing Li
author_facet Wei Liu
Xinyu Wang
Yating Chen
Jiapei Yuan
Huiyu Zhang
Xin Jin
Yuying Jiang
Junjing Cao
Zibin Wang
Shuo Yang
Bingwei Wang
Tinghe Wu
Jing Li
author_sort Wei Liu
collection DOAJ
description Abstract As functional derivatives of mesenchymal stem cells (MSCs), small extracellular vesicles (sEVs) have garnered significant attention and application in regenerative medicine. However, the technical limitations for large‐scale isolation of sEVs and their heterogeneous nature have added complexity to their applications. It remains unclear if the heterogeneous sEVs represent different aspects of MSCs functions. Here, we provide a method for the large‐scale production of sEVs subpopulations derived from human umbilical cord mesenchymal stem cells (HucMSCs), utilizing tangential flow filtration combined with size exclusion chromatography. The resulting subpopulations, S1‐sEVs and S2‐sEVs, exhibited stable variations in size, membrane‐marked proteins, and carrying cargos, thereby displaying distinct functions both in vitro and in animal disease models. S1‐sEVs, that highly expressed CD9, HRS and GPC1, demonstrated a greater immunomodulatory impact, while S2‐sEVs with enriched expression of CD63 and FLOT1/2 possessed enhanced capacities in promoting cell proliferation and angiogenesis. These discrepancies are attributed to the specific proteins and miRNAs they contain. Further investigation revealed that the two distinct sEVs subpopulations corresponded to different biological processes: the ESCRT pathway (S1‐sEVs) and the ESCRT‐independent pathway represented by lipid rafts (S2‐sEVs). Therefore, we propose the potential for large‐scale isolation and purification of sEVs subpopulations from HucMSCs with distinct functions. This approach may provide advantages for targeted therapeutic interventions in various MSC indications.
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spelling doaj-art-435a5c50bb0f4c31bf1aa050169d13ab2025-08-20T02:41:52ZengWileyJournal of Extracellular Vesicles2001-30782025-01-01141n/an/a10.1002/jev2.70029Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatographyWei Liu0Xinyu Wang1Yating Chen2Jiapei Yuan3Huiyu Zhang4Xin Jin5Yuying Jiang6Junjing Cao7Zibin Wang8Shuo Yang9Bingwei Wang10Tinghe Wu11Jing Li12State Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaWuxi Maternity and Child Health Care Hospital Affiliated Women's Hospital of Jiangnan University Wuxi ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaCenter for Analysis and Testing Nanjing Medical University Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaDepartment of Pharmacology Nanjing University of Chinese Medicine Nanjing ChinaKornelis Bio‐pharmaceutical Company Limited Nanjing ChinaState Key Laboratory of Reproductive Medicine and offspring health Nanjing Medical University Nanjing ChinaAbstract As functional derivatives of mesenchymal stem cells (MSCs), small extracellular vesicles (sEVs) have garnered significant attention and application in regenerative medicine. However, the technical limitations for large‐scale isolation of sEVs and their heterogeneous nature have added complexity to their applications. It remains unclear if the heterogeneous sEVs represent different aspects of MSCs functions. Here, we provide a method for the large‐scale production of sEVs subpopulations derived from human umbilical cord mesenchymal stem cells (HucMSCs), utilizing tangential flow filtration combined with size exclusion chromatography. The resulting subpopulations, S1‐sEVs and S2‐sEVs, exhibited stable variations in size, membrane‐marked proteins, and carrying cargos, thereby displaying distinct functions both in vitro and in animal disease models. S1‐sEVs, that highly expressed CD9, HRS and GPC1, demonstrated a greater immunomodulatory impact, while S2‐sEVs with enriched expression of CD63 and FLOT1/2 possessed enhanced capacities in promoting cell proliferation and angiogenesis. These discrepancies are attributed to the specific proteins and miRNAs they contain. Further investigation revealed that the two distinct sEVs subpopulations corresponded to different biological processes: the ESCRT pathway (S1‐sEVs) and the ESCRT‐independent pathway represented by lipid rafts (S2‐sEVs). Therefore, we propose the potential for large‐scale isolation and purification of sEVs subpopulations from HucMSCs with distinct functions. This approach may provide advantages for targeted therapeutic interventions in various MSC indications.https://doi.org/10.1002/jev2.70029mesenchymal stem cellssEVssize exclusion chromatographytangential flow filtrationtargeted therapies
spellingShingle Wei Liu
Xinyu Wang
Yating Chen
Jiapei Yuan
Huiyu Zhang
Xin Jin
Yuying Jiang
Junjing Cao
Zibin Wang
Shuo Yang
Bingwei Wang
Tinghe Wu
Jing Li
Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography
Journal of Extracellular Vesicles
mesenchymal stem cells
sEVs
size exclusion chromatography
tangential flow filtration
targeted therapies
title Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography
title_full Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography
title_fullStr Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography
title_full_unstemmed Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography
title_short Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography
title_sort distinct molecular properties and functions of small ev subpopulations isolated from human umbilical cord mscs using tangential flow filtration combined with size exclusion chromatography
topic mesenchymal stem cells
sEVs
size exclusion chromatography
tangential flow filtration
targeted therapies
url https://doi.org/10.1002/jev2.70029
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