Preparation method of human dendritic cell tumour vaccine

A technology of dendritic cells and tumor vaccines, applied in the field of preparation of human dendritic cell tumor vaccines, can solve the problems of low purity and maturity of DCs, improve the purity and maturity of DCs, facilitate treatment, and promote fusion Effect

Inactive Publication Date: 2015-04-08
深圳市赛欧细胞技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] Aiming at the problem of low purity and maturity of DCs in the preparation process of human dendritic cell tumor vaccines in the prior art, the present invention provides a preparation method of human dendritic cell tumor vaccines, which can increase the maturity of DCs and the purity is greatly improved

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  • Preparation method of human dendritic cell tumour vaccine

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Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1: Preparation of human dendritic cell tumor vaccine and verification experiment of DC maturation-promoting agent, including the following steps:

[0038] (1) Take 50 mL of donated human peripheral venous blood, use a blood cell separator to obtain peripheral blood mononuclear cells (PBMC), and the cell density is higher than 1×10 8 ~2×10 8 / mL;

[0039] (2) The resulting PBMCs were resuspended in a T175mL cell culture flask, and 10mL AMI-ⅴ serum-free medium and 10% autologous serum were added to adjust the cell density to 5×10 6 / mL at 37°C, 5% CO 2 Under the condition of , adhere to the wall and culture in the incubator for 2 hours;

[0040] (3) Gently shake the cell culture flask to remove the supernatant. Add 10mL AMI-ⅴ serum-free medium and 10% autologous serum again, repeat the above operation twice;

[0041] (4) Add 20 mL (0.2 to 1 times) of AMI-ⅴ serum-free medium and 5% autologous serum containing 100 ng / mL rhGM-CSF and 20 ng / mL rhIL-4 to the ab...

Embodiment 2

[0052] The preparation of human dendritic cell tumor vaccine comprises the following steps:

[0053] (1) Take 50 mL of donated human peripheral venous blood, use a blood cell separator to obtain peripheral blood mononuclear cells (PBMC), and the cell density is higher than 1×10 8 ~2×10 8 / mL;

[0054] (2) The resulting PBMCs were resuspended in a T175mL cell culture flask, and 10mL AMI-ⅴ serum-free medium and 10% autologous serum were added to adjust the cell density to 5×10 6 / mL at 37°C, 5% CO 2 Under the condition of , adhere to the wall and culture in the incubator for 2 hours;

[0055] (3) Gently shake the cell culture flask to remove the supernatant. Add 10mL AMI-ⅴ serum-free medium and 10% autologous serum again, repeat the above operation twice;

[0056] (4) Add 20 mL of AMI-ⅴ serum-free medium containing 100 ng / mL rhGM-CSF, 20 ng / mL rhIL-4 and 5% autologous serum to the above-mentioned cell culture flask, and continue to incubate at 37°C, 5% CO 2 Cultivate in th...

Embodiment 3

[0061] The DC maturation promoters were 1000U / mL TNF-α, 10ng / mL IL-6, 10ng / mL IL-1β, 0.15KE / mLOK-432, 8μg / mL gp100 280-288 , the preparation steps, processes and detection methods of other human dendritic cell tumor vaccines are the same as in Example 1.

[0062] The maturity test results are shown in Table 2:

[0063] Table 2 The detection results of mature human dendritic cells in Example 3

[0064] Test items

[0065] It can be found from Table 2 that mature DCs have a high degree of maturity and are very suitable for the preparation of DC tumor vaccines.

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Abstract

The invention provides a preparation method of a human dendritic cell tumour vaccine and belongs to the technical field of cell engineering. The human dendritic cell tumour vaccine is characterized in that a DC maturity acceleration agent is added in a process that the dendritic cell is matured from being unmatured, and the DC maturity acceleration agent comprises OK-432 and a gp100 combined DC maturity acceleration agent. By adopting the human dendritic cell tumour vaccine, cancer patients can be treated or high risk of people suffering from cancer can be avoided. The human dendritic cell tumour vaccine prepared by adopting the preparation method provided by the invention has the advantages that DC purity and maturity of the human dendritic cell tumour vaccine are greatly improved, fusion of DC and a tumour antigen is promoted, and treatment of cancer can be facilitated; meanwhile, a preparation technology is simple, and cost is low.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and in particular relates to a preparation method of a human dendritic cell tumor vaccine. Background technique [0002] The current conventional tumor treatment methods include surgical treatment, radiotherapy and chemotherapy, etc. The main problem faced by these treatment methods is the high rate of tumor recurrence and metastasis. The reason is that the tumor stem cells in the tumor are not all killed, which makes it easier for the cancer tissue to regenerate and re-form the tumor. Dendritic cells (DC, Dendritic Cell) are known to be the most powerful professional antigen-presenting cells in the human body, and have the function of activating T cells and killing tumor cells. Figuratively speaking, DC is equivalent to the messenger between tumor antigens and T cells, and can transmit information between the two parties. In particular, DCs sensitized with different forms of tumor ant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00
Inventor 崔兴日张涤平
Owner 深圳市赛欧细胞技术有限公司
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