Optimization Of Culture Dendirtic Cell In Vitro

1. Research background:The pillars of human cancer therapy have historically been surgery, radiotherapy, and chemotherapy, but a fourth modality of immunotherapy has been well documented which had benefited for inoperable cancers and other crude immunostimulants showed benefits that led to regulatory approval of their use in some solid tumors such as thyroid cancer and bladder cancer。The immunotherapy of cancer has made significant strides in the past few years due to improved understanding of the underlying principles of tumor biology and immunology. These principles have been critical in the development of immunotherapy in the laboratory and in the implementation of immunotherapy in the clinic.This improved understanding of immunotherapy, enhanced by increased insights into the mechanism of tumor immune response and its evasion by tumor.Also important, this improved understanding of immunotherapy and the mechanisms underlying immunity in cancer has fueled an expanding array of new therapeutic agents for a variety of cancers.GM-CSF is the central mediator for the generation of DC in vitro. Its major function appears to be to prolong cellular survival. It promotes the differentiation of monocytes to large macrophage-like cells, increasing their metabolism and function as APC by enhancing MHC Ⅱ expression. G-CSF is used to regulate the production of neutrophils and enhances their maturation. It is widely used for patients with neutropenia after chemotherapy. More recently, G-CSF has been shown to mobilize T helper2preferentially, inducing dendritic cells over those that induce T helper1cells.2. The purpose and significance:Currently, the treatment of autoimmune cell therapy as a new method, has been included in the category of third class medical technology clinical, my team has won the third class of clinical medical technology application permissions, and the early clinical treatment of immune cells work.This experiment by comparing two kinds of cytokines G-CSF and GM-CSF on DC cell proliferation, the effects of cytokines and preliminary discussion on the role of DC differentiation and mature, and by adding different concentrations of autologous serum, analysis in vitro induced to generate the number of DC, surface markers and stimulate the function of lymphocyte proliferation, explore the best conditions of peripheral blood DC in vitro, in order to optimize the DC in vitro cultivation scheme. For future clinical treatment to provide both quantity and quality qualified DC cells and lay a foundation for science of cellular immune treatment personalized solutions.3. Method:Respectively using g-csf and gm-csf induced peripheral blood mononuclear cell differentiation in vitro DC; Adding different concentration of autologous serum culture DC.4. Result:(1) G-CSF and IL-4combination, GM-CSF and IL-4combination, can be induced in vitro PBMC differentiate into DC.(2) GM-CSF and IL-4combination of DC induced on the quantity and purity were superior to G-CSF combined with IL-4;(3) measured by MLR mixed lymphocyte reaction two combination differentiation of DC in stimulating the role of T cell proliferation much;(4) in vitro cultivation of DC, add10%of autologous serum appropriately, can improve the yield of DC.