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CIK cell culture method

A cell culture and cell technology, applied in the direction of cell culture active agent, culture process, tissue culture, etc., can solve the problems of poor activity and proliferation ability of CIK cells, to improve activity and proliferation ability, adequate nutrition and good environment, shorten cycle effect

Inactive Publication Date: 2017-09-08
DONGGUAN BOALAI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, most of the CIK cells cultured by the existing CIK cell method have poor activity and proliferation ability.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] In this embodiment, a CIK cell culture method comprises the following steps:

[0042] (1) Resuspend peripheral blood mononuclear cells in CIK medium to obtain cell suspension;

[0043] (2) Add the cell suspension to the T175 culture flask that has been pre-coated with anti-CD3 monoclonal antibody and retronectin, and then place it at 37°C, 5% CO 2 In a saturated humidity incubator; adjust the cell density to 7×10 before culturing 5-2×10 6 / mL;

[0044] (3) On the third day of culture, 60 mL of CIK culture medium was supplemented;

[0045] (4) On the 6th day of culture, after adding the CIK culture solution, pour the original culture solution in the original T175 culture bottle into a new culture bottle, blow off the cells on the wall of the original T175 culture bottle with the culture solution, and pour Put into a new culture bottle, add CIK culture solution to a total volume of 400mL, divide the above culture solution into four culture bags, add CIK culture solut...

Embodiment 2

[0064] In this embodiment, a CIK cell culture method comprises the following steps:

[0065] (1) Resuspend peripheral blood mononuclear cells in CIK medium to obtain cell suspension;

[0066] (2) Add the cell suspension to the T175 culture flask that has been pre-coated with anti-CD3 monoclonal antibody and retronectin, and then place it at 37°C, 5% CO 2 In a saturated humidity incubator; adjust the cell density to 7×10 before culturing 5 -1.5×10 6 / mL;

[0067] (3) On the third day of culture, 60 mL of CIK culture medium was supplemented;

[0068] (4) On the 6th day of culture, after adding the CIK culture solution, pour the original culture solution in the original T175 culture bottle into a new culture bottle, blow off the cells on the wall of the original T175 culture bottle with the culture solution, and pour Put into a new culture bottle, add CIK culture solution to a total volume of 400mL, divide the above culture solution into four culture bags, add CIK culture so...

Embodiment 3

[0088] In this embodiment, a CIK cell culture method comprises the following steps:

[0089] (1) Resuspend peripheral blood mononuclear cells in CIK medium to obtain cell suspension;

[0090] (2) Add the cell suspension to the T175 culture flask that has been pre-coated with anti-CD3 monoclonal antibody and retronectin, and then place it at 37°C, 5% CO 2 In a saturated humidity incubator; adjust the cell density to 7×10 before culturing 5 -1.5×10 6 / mL;

[0091] (3) On the third day of culture, 60 mL of CIK culture medium was supplemented;

[0092] (4) On the 6th day of culture, after adding the CIK culture solution, pour the original culture solution in the original T175 culture bottle into a new culture bottle, blow off the cells on the wall of the original T175 culture bottle with the culture solution, and pour Put into a new culture bottle, add CIK culture solution to a total volume of 400mL, divide the above culture solution into four culture bags, add CIK culture solu...

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Abstract

The invention relates to the biotechnology field and in particular to a CIK cell culture method. The method comprises the following steps: (1) heavy-suspending a mononuclear cell of peripheral blood in CIK culture solution, to obtain cell suspension; (2) adding the cell suspension into a culture bottle pre-enveloped by an anti-CD3 monoclonal antibody and retronectin, culturing in a culture box with 5% of CO2 saturation humidity in 37 DEG C of the temperature; (3) replenishing the CIK culture solution in the third day of the culturing; (4) in the sixth day of the culturing, after replenishing the CIK culture solution, executing the culture expansion; (5) replenishing the CIK culture solution in the ninth day of the culturing; and (6) culturing for 13-14 days, and harvesting the cells. The method is capable of remarkably improving the proliferation rate of the immune cell, especially the proliferation rate of CD3+CD56+ effector cell, and improving the killing activity thereof.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for culturing CIK cells. Background technique [0002] Cell adoptive immunotherapy is a biological treatment method in which immune cells induced in vitro with strong tumoricidal activity are returned to patients. It not only has the ability to kill residual cancer cells in the body, but also enhances the immunity of the host. One of the important means of biological therapy. [0003] Cytokine induced killer cells (CIK) are a group of heterogeneous cells obtained by co-culturing and stimulating human peripheral blood mononuclear cells (PBMCs) with various cytokines. Effector cells are CD3 + CD56 + Double-positive cells have both the strong anti-tumor activity of T lymphocytes and the characteristics of non-major histocompatibility complex (MHC)-restricted tumor killing. CIK cells have the characteristics of fast proliferation, high tumor killing activity, broad tumor ki...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N2500/50C12N2501/2302C12N2501/24C12N2501/515C12N2501/90C12N2501/91
Inventor 罗天恩
Owner DONGGUAN BOALAI BIOLOGICAL TECH CO LTD
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