HucMSC-Exo driving neutrophil to N2 promoting angiogenic phenotypic transformation
Background Human umbilical cord mesenchymal stem cell-derived exosomes(HucMSC-Exo)have become a promising therapeutic tool in the field of regenerative medicine,but the exact mechanism of their action is still poorly understood.Objective To explore whether HucMSC-Exo could drive neutrophil switching to N2 proangiogenic subtype.Methods HucMSC-Exo was isolated and purified by ultracentrifugation,and the isolated exosomes were characterized by transmission electron microscopy,particle size analysis and western blot.Isolated HucMSC-Exo was co-cultured with neutrophils in vitro and the expression of phenotype-related molecules in neutrophils was detected using qRT-PCR.After co-culture with HucMSC-Exo,neutrophils were subjected to cell-cell co-culture system with vascular endothelial cells,and the effects of neutrophils on the proliferation,migration and invasion ability of vascular endothelial cells were observed by CCK-8 assay,Edu assay,scratch assay and Transwell assay,respectively.Gene expression changes related to angiogenesis in neutrophils after receiving HucMSC-Exo stimulation were analyzed using qRT-PCR.Results The extracts were shown in transmission electron microscopy as a round-like membrane-like structure with a diameter of approximately 100 nm,and were able to express the exosomal markers CD9 and Alix,indicating that the extracts were HucMSC-Exo.Compared to the PBS-treated group,treatment of neutrophils with HucMSC-Exo revealed significantly increased expression of N2 markers,such as Arg1(P<0.05),CD163(P<0.05)and CD206(P<0.05)by qRT-PCR.Following co-culture of HucMSC-Exo-stimulated neutrophils with vascular endothelial cells,the vascular endothelial cells became more proliferative,migratory,and invasive as observed by CCK-8 assay,Edu assay,scratch assay,and Transwell assay,respectively(P<0.05).Compared with the PBS-treated group,neutrophils in HucMSC-Exo-treated group displayed elevated expression of pro-angiogenic factors BV8(P<0.05)and VEGFα(P<0.05).Conclusion HucMSC-Exo converts neutrophils to the N2 phenotype,and this N2 type of neutrophil promotes vascular endothelial cell proliferation,migration,and invasion.
human umbilical cord mesenchymal stem cellsexosomesneutrophilsphenotype switchingangiogenesis
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