The Effects Of HucMSCs-derived Exosome On Phenotype And Function Of Mononuclear Macrophage

Objective: To investigate the effects of human umbilical cord mesenchymal stem cells and exosome on phenotype and function of mononuclear macrophages in vitro.Methods: 1.The huc MSCs were isolated and cultivated from human umbilical cord by tissue adherent in vitro,and the 3~6 generation huc MSCs were used for subsequent experiments.2.To explore the co-culture conditions of the huc MSCs and mononuclear macrophages,then the luminex method is used to screen the secretory changes of inflammatory factors.Further more,some relevant cytokines were detected.3.Exosome was separated and concentrated by ultracentrifugation from the huc MSCs.And exosome was identified by western blot and electron microscopy.Then the concentration of exosome was measured by BCA protein assay and adjusted to 0.1 mg/m L of spare.4.After mononuclear macrophages were treated with huc MSCs or huc MSCs derived exosome,the expression of IL-8?IL-12?IL-10 m RNA were assessed by general PCR and Real-time fluorescence quantitative PCR(q RT-PCR)Cell proliferation.The cytokines of IL-8?IL-12?IL-10 proteins from mononuclear macrophages and the co-culture system were explored by western blot and ELISA assay.The proliferation was detected by MTT assay,cellular apoptosis was measured by Annexin V-FITC and PI double stain,and the cell migration function was tested by transwell assay.Results: 1.Human umbilical cord mesenchymal stem cells(hucMSCs)and their source of exosome could be successfully isolated and cultured.2.When the ratio of the huc MSCs to mononuclear macrophages was 1: 2,both the cells growth were well.And after cultivating 24 h,the changes of the secretion level of the related inflammatory factors were relatively obvious.3.In some way,the huc MSCs and exosome could suppress the expression of cytokine IL-8 ? IL-12 of the M1 macrophages and up-regulate the expression of cytokine IL-10 of M2 macrophages.4.After treated with huc MSCs or exosome on mononuclear macrophages,the proliferation and migration were promoted and the apoptosis was inhibited in vitro.Conclusions: 1.In a certain extent,the hucMSCs and their exosome can promote the polarization of mononuclear macrophages to M2 macrophages in vitro.2.The huc MSCs and their exosome can promote the proliferation,migration and inhibit the apoptosis in mononuclear macrophages.3.Exosome is likely to be one of the mechanisms of huc MSCs to play a role in phenotype and function of mononuclear macrophages in vitro.