Comprehensive proteomic analysis of exosome mimetic vesicles and exosomes derived from human umbilical cord mesenchymal stem cells

Abstract

Background 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. Methods 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. Results 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. Conclusions 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.

Engineered Nanovesicles from Fibroblasts Modulate Dermal Papillae Cells In Vitro and Promote Human Hair Follicle Growth Ex Vivo

“…Therefore, we confirmed that three of the classical EV protein markers (Alix, HSP90, and Flotillin-1), often used to characterize EVs [ 20 , 21 ], were expressed in FB-eNVs. In addition, cellular proteins such as GM130 and calnexin are present in FB-eNVs as the FB-eNVs are synthesized through direct cell extrusion and may contain all organelle components [ 22 ], including the Golgi apparatus and ER. Furthermore, biological cargoes (including proteins) are randomly packed into eNVs, rather than the selective packing of specific proteins in EVs [ 23 , 24 , 25 ].…”

“…Furthermore, biological cargoes (including proteins) are randomly packed into eNVs, rather than the selective packing of specific proteins in EVs [ 23 , 24 , 25 ]. Nonetheless, EVs and eNVs (also called EV-mimetics) have been demonstrated to have 80% similar proteomes [ 22 ]. Next, we confirmed that the generated FB-eNVs were intact/round and approximately 100 nm in diameter, and within the range of typical EVs (e.g., exosomes) [ 3 , 26 ].…”

The roles of mesenchymal stem cell-derived exosomes in diabetes mellitus and its related complications

“…Diabetes-related cognitive impairment is a global challenge, with epidemiological studies reporting that the incidence of dementia in diabetic patients is two to three times higher than in nondiabetic patients. Multiple mechanisms are thought to contribute to diabetes-related dementia, including abnormal glucose metabolism (e.g., hyperglycemia and hypoglycemia), abnormal insulin action (e.g., insulin deficiency and insulin resistance), vascular abnormalities, and oxidative stress in the central nervous system (86,135). The damage to hippocampal neurons and astrocytes is considered the most critical problem of cognitive impairment caused by diabetes.…”

“…Proteomic analysis of EMVs and exosomes derived from HucMSCs revealed that they share about 80% of the same protein while also containing unique proteins. EMVs and exosomes derived from HucMSCs have tissue repair ability, promoting wound healing and angiogenesis in mice (135). Future research must determine whether different MSCderived EMVs have the same therapeutic effect as exosomes and whether they can be applied to diabetes and associated complications and other diseases.…”

Chondrogenic Potential of Human Umbilical Cord Mesenchymal Stem Cells Cultured with Exosome-Depleted Fetal Bovine Serum in an Osteoarthritis Mouse Model

“…However, above all, the exosomes in FBS may affect the proliferation and differentiation capability of HUCMSCs. The HUCMSCs can secrete exosomes to affect many cell types in the microenvironment [ 14 , 15 ]. We expected the benefit of Exo(-)FBS on the differentiation of HUCMSCs, including no influence on HUCMSCs’ exosome secretion and maximizing self-secreted exosomes’ functionality, which may enhance the differentiation capability of HUCMSCs.…”