Method for preparing dendritic cell vaccine

Abstract

The invention relates to the technical field of biology, in particular to a method for preparing dendritic cell vaccine. The method is characterized by comprising the following preparation steps: (1) taking autologous blood of a patient, and separating red blood cells, mononuclear cells and plasma from separating medium of human lymphocyte; (2) adjusting concentration of the mononuclear cells; (3) adding the solution into a six-pore plate and performing adherence for 16 hours; (4) sucking and discharging non-adherent cells on the upper layer of each pore of the six-pore plate; (5) adding 3ml of dendritic cell (DC) culture medium, placing the solution in a carbon dioxide cultivating box with the concentration of carbon dioxide of 5% at the temperature of 37 DEG C to perform cultivation for 2-3 days; (6) performing half-quantity liquid changing; (7) enabling the concentration of the autologous tumor antigen in each pore of the six-pore plate to be 20 mu g / ml by loading the autologous tumor antigen in the sixth day; and (8) detecting the quantity and maturity of the matured DC cells after 24 hours. Compared with the prior art, the number of the matured DC cells at least reaches 1*107, and the maturity is larger than 85%.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method of a dendritic cell vaccine. Background technique [0002] DCs are called dendritic cells, which are important immune cells in the body and have broad application prospects in cellular immunotherapy. Under the stimulation of specific tumor antigens, mature dendritic cells can express a variety of cytokines and chemotactic Factors, such as MHC Ⅱ molecule HLA-DR, co-stimulatory molecules CD80, CD86 and other cytokines, can effectively induce antigen-specific and non-specific immune responses and enhance the immune function of the body. However, the proportion of DC in human peripheral blood is very low, so increasing the expansion multiple and increasing the number of mature dendritic cells play a key role in the therapeutic effect. [0003] Dendritic cells are very hot now. The conventional technique is to use GM-CSF+IL4. The amplification factor of this method i...

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Problems solved by technology

[0003] Dendritic cells are very hot now. The conventional technique is to use GM-CSF+IL4. The amplification factor of this method is not high, and the maturity is not high. Generally, the number of mature dendritic cells can only reach 2×10 6 , the purity of mature dendritic cells is 60%, see figure 1

There is also the method of adding Flt3 for amplification, but chemical drugs are also added, and the amplification factor and maturity are not high

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