Immune cell capable of preventing tumorigenesis as well as preparation method and application thereof

An immune cell and tumor technology, applied in the field of immune cells, can solve the problems of difficult cell-derived treatment side effects, enhanced killing cytotoxic activity, undisclosed tumor prevention, etc., to stimulate T cell proliferation, induce tumor regression, and improve cell The effect of multiplication factor

Active Publication Date: 2022-03-01
杭州中赢生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In international tumor immunotherapy, lymphokine-activated killer cells (LAK cells), anti-CD3 monoclonal antibody-induced killer cells, and tumor-infiltrating lymphocytes (TIL cells) have been experienced successively. The source is difficult (such as TIL) or the side effects of treatment are relatively large (such as LAK), so there are fewer and fewer studies and reports
[0005] CN 102321577 B discloses a method for preparing anti-tumor adoptive immune cells and the prepared immune cells. The method includes collecting and separating peripheral blood mononuclear cells, adding cytokines to induce culture to obtain killer cells CIK, and the cytokines include CD28McAb. Preparation The immune cells can increase the expansion of effector cells and enhance the killing cytotoxic activity. Although the obtained DC-CIK becomes tumor-specific cytotoxic cells and improves the overall tumor killing effect of the CIK population, it does not disclose the effect of preventing effect on tumorigenesis

Method used

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  • Immune cell capable of preventing tumorigenesis as well as preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0029] SD rats purchased from the Experimental Animal Center of Wenzhou Medical University were raised in the SPF environment of the animal room, and allowed to obtain feed and water arbitrarily, and acclimated to the environment for 1 week. All principles and procedures of animal experiments were in compliance with the relevant provisions of the "Regulations on the Administration of Experimental Animals".

[0030] The rats were numbered, and then the peripheral blood of the rats was extracted sequentially under sterile conditions, added to a centrifuge tube with heparin saline, shaken gently, fully anticoagulated, diluted 1:1 with normal saline, and added to the lymphocyte separation medium , centrifuge at 2000r / min for 15min at room temperature, suck out the PBMCs and transfer them into a new centrifuge tube, add physiological saline, mix them gently with a pipette, centrifuge at 2000r / min for 15min, discard the supernatant, add AIM-V (GIBCO) for serum-free culture Base adju...

Embodiment 2

[0032] Dissolve one tube of 2.5 mg Retronectin in 250 mL of normal saline in advance to make a coating solution with a final concentration of 10 μg / ml. Put the coating solution in a culture bottle and let it stand overnight. The next day, the coating solution was discarded, and the culture flask was washed twice with physiological saline to obtain a culture flask coated with Retronectin.

[0033] The non-adherent cells after adherent culture in Example 1 were resuspended in 10 mL of AIM-V medium, transferred into culture flasks coated with Retronectin, added cytokine IFN-γ 1000 μg / ml, at 37°C, 5% CO 2 Cultured in a culture environment, the cell density was controlled at 1-5×10 during the culture process. 7 / mL.

[0034] On the second day, add cytokines CD3McAb 2 μg / mL, CD28McAb 1 μg / mL, and IL-2 1000 μg / mL to the culture flask, at 37°C, 5% CO 2 Cultured in a culture environment, the cell density was controlled at 1-5×10 during the culture process. 7 / mL;

[0035] On the ...

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Abstract

The invention discloses an immune cell capable of preventing tumorigenesis. The invention discloses a preparation method of the immune cell, which comprises the following steps: placing a peripheral blood mononuclear cell and a culture medium in a culture bottle coated with Retronectin, and adding 500-2000 [mu] g / ml of a cell factor IFN-gamma for culture; culturing to the second day, adding 0.5-5 [mu] g / mL of cell factors CD3McAb, 0.5-2 [mu] g / mL of CD28McAb and 500-2000 [mu] g / mL of IL-2, and continuing culturing; culturing to the 7th day, adding 0.1-0.5 [mu] g / mL of cell factors CD-40 L and 0.1-0.5 [mu] g / mL of GM-CSF, and continuing culturing; the culture medium is supplemented every 3-4 days in the culture process, 500-2000 [mu] g / mL of cytokine IL-2 is supplemented every 1-2 days, and the cells are continuously cultured for 14 days until the cells are mature.

Description

technical field [0001] The invention relates to the technical field of immune cells, in particular to an immune cell capable of preventing tumor occurrence, its preparation method and application. Background technique [0002] Tumor is a common and frequently-occurring disease that seriously threatens human health. In my country, there are more than 1.6 million malignant tumor patients every year, and cancer is surpassing cardiovascular disease as the first cause of death. Tumor biological therapy is the fourth largest treatment method after surgery, radiotherapy, and chemotherapy. The development of chemotherapy has entered a plateau, and people are paying more and more attention to the prevention of tumors. [0003] If the rising incidence of cancer could be stopped, reduced, or even prevented, it would be beneficial to cancer patients and healthcare providers around the world by avoiding the need for expensive treatment regimens and intense medications, so whether it is f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A61K35/17A61P35/00A61P35/04
CPCC12N5/0638A61K35/17A61P35/00A61P35/04C12N2533/52C12N2501/24C12N2501/515C12N2501/51C12N2501/2302C12N2501/52C12N2501/22C12N2509/00C12N2500/90Y02A50/30
Inventor 不公告发明人
Owner 杭州中赢生物医疗科技有限公司
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