Adipose-derived Stem Cells In Inflammatory Microenvironment Inhibit IFN-? And TNF-? Secretion In Peripheral Blood Mononuclear Cells

Background: The repair of skin tissue defects caused by various factors such as wounds,diabetic foot,and other bacterial infections has been a difficult problem for plastic surgery and other surgeries.Adipose-derived stem cells(ADSCs)are a type of pluripotent mesenchymal cells that have the ability to differentiate into adipocytes,osteoblasts,and chondrocytes.More and more recent studies show that autologous or allogeneic ADSCs have the unique ability to play a regulatory role in adaptive and innate immunity.In recent years,a large number of studies have found that ADSCs can regulate immune cells by distant regulation of immune cells through migration to inflammatory sites or secretion of cytokines and other mechanisms.Peripheral blood mononuclear cells(PBMC),including lymphocytes,monocytes,macrophages,and dendritic cells,have always played an important role in immune inflammation and other reactions.Phytohemagglutinin(PHA)is a complex of protein and oligosaccharides.As a stimulating factor,it can activate lymphocytes in PBMC and then divide and proliferate,release lymphokines,and increase Phagocytic phagocytosis mimics the PBMC's microenvironmental state during inflammation.This experiment is to explore the regulation of ADSCs on the ability of PBMCs to secrete inflammatory factors in the inflammatory microenvironment.Objective: Extract and identify ADSC and PBMC,stimulate PBMC with PHA to simulate the inflammatory response microenvironment,co-culture ADSCs pretreated with inflammatory factors and PBMC,explore the effect of ADSC on the ability of PBMC to secrete inflammatory factors,and then explore ADSCs on the inflammatory microenvironment role.Methods: 1.Extraction,culture and identification of ADSCs and PBMCs;2.Activate PBMCs with PHA and check the results after activation;3.Interferon-?(IFN-?)and Tumor Necrosis Factor-?(TNF-?)pre-stimulate adipose-derived stem cells for 24 hours to activate their immunoregulatory capacity.PHA-activated PBMCs are co-cultured with pre-stimulated ADSCs.After co-culture for 24 hours,the supernatantslevel of IFN-? and TNF-? are detected;4.Use SPSS 19.0 and Graph pyramid 5software to analyze the data,P <0.05 was considered statistically significant.Results: 1.Successful extraction of human adipose stem cells and peripheral blood mononuclear cells;2.PHA can effectively activate PBMC and promote the secretion of more pro-inflammatory cytokines IFN-?and TNF-?;3,IFN-?and TNF-?pre-treated ADSCs can effectively inhibit PHA-activated PBMCs from secreting IFN-?and TNF-?.Conclusion: 1.Confirm that PHA can stimulate PBMC to secrete more IFN-?and TNF-?;2.ADSCs pre-stimulated by inflammatory factors can significantly inhibit the ability of PBMCs to secrete IFN-?and TNF-?after PHA stimulation;3.It was confirmed that ADSCs can regulate the inflammatory response.