Method for preparing dendritic cell vaccine

A technology for dendritic cell and vaccine preparation, which is applied in the field of dendritic cell vaccine preparation, can solve the problems of low maturity, low amplification multiple and maturity, and low amplification multiple, and achieves high purity, Enhance the amplification factor and the effect of strong specific lethality

Inactive Publication Date: 2013-01-02
FUDAN UNIV
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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 ca...

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  • Method for preparing dendritic cell vaccine
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Embodiment Construction

[0009] Below in conjunction with example the present invention will be further described.

[0010] (1) Take 18ml of the patient's autologous blood, and take 1ml of the patient's autologous blood for flow cytometry, which is used for the evaluation of immune indicators before treatment, and the other 17ml of the patient's autologous blood is separated from red blood cells and monocytes Lymphocytes, plasma;

[0011] (2) Use GT-T551 serum-free medium to adjust the concentration of mononuclear lymphocytes to 2×10 6 a / ml;

[0012] (3) Add the adjusted concentration of mononuclear lymphocytes into the 6-well plate at 3ml / well, and adhere to the wall for 16 hours;

[0013] (4) Discard the upper layer of non-adherent cells in each well of the 6-well plate, leaving the lower layer of adherent immature DC cells in each well;

[0014] (5) Add 3ml of DC medium to each well of the 6-well plate and place it in a carbon dioxide incubator, where CO 2 The concentration is 5%, the temperatu...

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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...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00C12N5/0784
Inventor 储以微郑秀娟张丹
Owner FUDAN UNIV
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