Exosomes derived from mesenchymal stem cells (MSCs) have shown to have effective application prospects in the medical field, but exosome yield is very low. The production of exosome mimetic vesicles (EMVs) by continuous cell extrusion leads to more EMVs than exosomes, but whether the protein compositions of MSC-derived EMVs (MSC-EMVs) and exosomes (MSC-exosomes) are substantially different remains unknown. The purpose of this study was to conduct a comprehensive proteomic analysis of MSC-EMVs and MSC-exosomes and to simply explore the effects of exosomes and EMVs on wound healing ability. This study provides a theoretical basis for the application of EMVs and exosomes. In this study, EMVs from human umbilical cord MSCs (hUC MSCs) were isolated by continuous extrusion, and exosomes were identified after hUC MSC ultracentrifugation. A proteomic analysis was performed, and 2315 proteins were identified. The effects of EMVs and exosomes on the proliferation, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) were evaluated by cell counting kit-8, scratch wound, transwell and tubule formation assays. A mouse mode was used to evaluate the effects of EMVs and exosomes on wound healing. Bioinformatics analyses revealed that 1669 proteins in both hUC MSC-EMVs and hUC MSC-exosomes play roles in retrograde vesicle-mediated transport and vesicle budding from the membrane. The 382 proteins unique to exosomes participate in extracellular matrix organization and extracellular structural organization, and the 264 proteins unique to EMVs target the cell membrane. EMVs and exosomes can promote wound healing and angiogenesis in mice and promote the proliferation, migration and angiogenesis of HUVECs. This study presents a comprehensive proteomic analysis of hUC MSC-derived exosomes and EMVs generated by different methods. The tissue repair function of EMVs and exosomes was herein verified by wound healing experiments, and these results reveal their potential applications in different fields based on analyses of their shared and unique proteins.

中文翻译：

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人脐带间充质干细胞衍生的外泌体模拟囊泡和外泌体的综合蛋白质组学分析

间充质干细胞（MSCs）衍生的外泌体已被证明在医学领域具有有效的应用前景，但外泌体的产量非常低。通过连续细胞挤压产生的外泌体模拟囊泡（EMV）比外泌体产生更多的EMV，但MSC衍生的EMV（MSC-EMV）和外泌体（MSC-外泌体）的蛋白质组成是否存在显着差异仍然未知。本研究的目的是对 MSC-EMV 和 MSC-外泌体进行全面的蛋白质组学分析，并简单探索外泌体和 EMV 对伤口愈合能力的影响。该研究为EMVs和外泌体的应用提供了理论依据。在本研究中，通过连续挤压分离人脐带间充质干细胞（hUC MSCs）的 EMV，并在 hUC MSC 超速离心后鉴定外泌体。进行了蛋白质组学分析，鉴定了 2315 种蛋白质。通过细胞计数 kit-8、划痕、transwell 和小管形成试验评估 EMV 和外泌体对人脐静脉内皮细胞 (HUVEC) 增殖、迁移和血管生成的影响。使用小鼠模式评估 EMV 和外泌体对伤口愈合的影响。生物信息学分析表明，hUC MSC-EMV 和 hUC MSC-外泌体中的 1669 种蛋白质在逆行囊泡介导的转运和囊泡从膜出芽中发挥作用。外泌体特有的 382 种蛋白质参与细胞外基质组织和细胞外结构组织，EMV 特有的 264 种蛋白质靶向细胞膜。EMVs和外泌体可以促进小鼠伤口愈合和血管生成并促进增殖，HUVECs的迁移和血管生成。本研究对不同方法产生的 hUC MSC 衍生的外泌体和 EMV 进行了全面的蛋白质组学分析。本文通过伤口愈合实验验证了 EMV 和外泌体的组织修复功能，这些结果基于对其共有和独特蛋白质的分析，揭示了它们在不同领域的潜在应用。