Protective effect of human umbilical cord mesenchymal stem cell-derived exosomes on vascular endothelial cells injured by hypoxia/reoxygenation
Objective To study the protective effect of human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSCs-exo)on vascular endothelial cell injured by hypoxia/reoxygenation(H/R).Methods Human umbilical cord mesenchymal stem cells(hUCMSCs)were extracted from the Wharton's jelly of full-term newborn umbilical cord by tissue adherence method.The surface markers CD34,CD45,CD73,CD90,CD105 of hUCMSCs were detected by flow cytometry.The differentiation ability of hUCMSCs was detected by adipogenic and osteogenic experiments.The exosomes in the culture supernatant of hUCMSCs were extracted by ultracentrifugation.The morphology of hUCMSCs-exo was detected by electron microscope and the marker proteins CD9 and CD63 of hUCMSCs-exo were detected by Western blot.HUVECs were cultured for 12 h with hypoxia(1％O2)and then incubated with normoxic(95％空气)for 12 h to establish an H/R model.Experimental groups were divided into control,H/R,hUCMSCs-exo groups.Typan blue staining was used to detect the cell viability of HUVECs,CCK-8 was used to detect the proliferation ability of HUVECs,Transwell assay was used to detect the migration ability of HUVECs,and tube formation assay was used to detect the angiogenic ability of HUVECs.Results The CD73,CD90 and CD105 markers of hUCMSCs extracted from Wharton's jelly were positive,but negative for CD34 and CD45 markers,and hUCMSCs had the adipogenic and osteogenic ability.The vesicle morphological characteristics of exosomes were observed by electron microscope,and the positive expressions of CD9 and CD63 were detected by Western blot.The cell viability and ability of proliferation and migration and angiogenesis were lower in H/R group than in control group.hUCMSCs-exo improved the cell viability of HUVECs after H/R,ameliorated their proliferation ability,migration ability and tube formation ability.Conclusion hUCMSCs-exo has a protective effect on the damaged HUVECs by H/R injury.
human umbilical cord mesenchymal stem cellexosomevascularendothelial cellhypoxia/reoxygen-ation
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