The Studies On Proliferation Culture Of DC In Vitro And The Effect Of Natural Medicine Preparation On DC

【Objective】Dendritic cells appear to be the strongest class of professional antigen-presenting cell.The present researches indicated that DCs can induce the tumor host produce specificity immune response.It enhances cytotoxin activeness of tumor host immunological reactivity cells.It has a high place on the immunity treatment of the tumor and the virus infection.In this study,we try to establish a stable method which can induce DC differentiation in vitro,The effect of natural drugs on DC's differentiation and mature could be absorbed with the different doses of natural medicine to be added on different phase of DC growth..The goal is to set up the foundation for the natural medicine screening of DC.【Methods】Peripheral blood mononuclear cell(PBMC)isolated from human peripheral blood by density gradient centrifugation were cultured.After incubation of PBMNC for 4 hours,DC was induced from adherent cells in Combination with Cytokine The biological characteristics of Derived DC were analyzed by flow cytometer(FACS),Scanning electron microscope and mixed lymphocyte reaction(MLR) in succession.A stable method which induce differentiation of DC in vitro was established.Astragelus polysaccharide and Lycium barbarum polysaccharide was added into the system in the growth process of DC along these lines.The effect to DC was analyzed by flow cytometer(FACS),Scanning electron microscope and mixed lymphocyte reaction(MLR).【Results】The monocytes isolated from human peripheral blood can be induced to mature DC with different cell factors in vitro.Observation with inverted microscope showed that the cells have the typical appearance after several day's cultivation.The data from flow cytometry and the MTT assay showed the remarkable difference compared with the blank group.The phenotypes of cells have the remarkable change by adding Astragelus polysaccharide and Lycium barbarum polysaccharide.Especially,Astragelus polysaccharide showed obvious effects on all the process of DC.The results of inverted microscope,flow cytometry and MTT assay all showed the remarkable difference compared with the blank group.【Conclusions】The study indicated that monocytes isolated from human peripheral blood could be induced to mature DC and proliferation with different cell factors in vitro.MTT assay showed that the matural DC which cultured in vitro have the strong capacity for stimulation of T cell sharply to be increased and tumour cells to be killed effectively,certain cell factors could be substituted by Astragalus and Lycium in induceing the peripheral blood monocytes to differentiate into functional DCs committedly.The anti-tumor treatment clinical practice of immune cells could be even much safely and economic in this way.The studies on Proliferation of DC and it's related natural medicine screening is worthed for further studying.