Co-culture method of DC cell and CIK cell

A cell culture and co-cultivation technology, applied in the field of cell culture, can solve the problems affecting the efficiency of DC cell antigen presentation, short survival period, slow maturation, etc., to improve the proliferation speed and killing activity, prolong the survival period, and increase the number Effect

Inactive Publication Date: 2015-03-25
山东世博金都药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the existing DC-CIK co-cultivation, the harvested DC cells are added to the CIK culture system, so that the antigen information carried by the DC cells can be extracted to the CIK cells through the combination of DC cells and CIK cells; but the culture of CIK The environment is not conducive to the growth of DC cells, and cannot effectively maintain the activity of

Method used

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  • Co-culture method of DC cell and CIK cell
  • Co-culture method of DC cell and CIK cell
  • Co-culture method of DC cell and CIK cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiments 1 to 4 are specific embodiments of the weaving method of a tubular fabric of the present invention, wherein embodiment 1 is the best embodiment.

[0049] Example 1

[0050] 1) Extract cells, the specific operation is as follows:

[0051] 1.1) Transfer peripheral blood: Collect 70ml of peripheral blood from the patient and transfer it to two 50ml centrifuge tubes, with an average of 35ml in each tube;

[0052] 1.2) Adding samples to the layer: Mix the blood sample and normal saline at a volume ratio of 1:1, and slowly add it to a 50ml sterile centrifuge tube filled with 20ml human lymphocyte separation medium at room temperature. The method is as follows: use a 10ml pipette to draw the blood sample, extend it to 0.5 cm above the surface of the human lymphocyte separation medium, let the first drop of blood sample slide down the tube wall slowly and naturally onto the surface of the human lymphocyte separation medium, and then Slowly rotate the centrifuge ...

Embodiment 2

[0074] 1) Extract cells, the specific operation is as follows:

[0075] 1.1) Transfer peripheral blood: Collect 50ml of peripheral blood from the patient and transfer it to two 50ml centrifuge tubes, with an average of 25ml per tube;

[0076] 1.2) Adding samples to the layer: mix the blood sample and normal saline at a volume ratio of 1:1, and slowly add it to a 50ml sterile centrifuge tube filled with 15ml human lymphocyte separation medium at room temperature. The method is as follows: use a 10ml pipette to draw the blood sample, extend it to 0.5cm above the liquid surface of the human lymphocyte separation medium, let the first drop of blood sample slide slowly and naturally along the wall of the tube to spread on the liquid surface of the human lymphocyte separation medium, and then Slowly rotate the centrifuge tube at a uniform speed, and at the same time slowly add the blood sample at a uniform speed, after balancing, centrifuge at 500 g, 30 min, RT, slowly rise and fall...

Embodiment 3

[0098] 1) Extract cells, the specific operation is as follows:

[0099] 1.1) Transfer peripheral blood: Collect 78ml of peripheral blood from the patient and transfer it to three 50ml centrifuge tubes, with an average of 26ml in each tube.

[0100] 1.2) Adding samples to the layer: mix the blood sample and normal saline at a volume ratio of 1:1, and slowly add it to a 50ml sterile centrifuge tube filled with 10ml human lymphocyte separation medium at room temperature. The method is as follows: use a 10ml pipette to draw the blood sample, extend it to 0.5 cm above the surface of the human lymphocyte separation medium, let the first drop of blood sample slide down the tube wall slowly and naturally onto the surface of the human lymphocyte separation medium, and then Slowly rotate the centrifuge tube at a uniform speed, and at the same time slowly add the blood sample at a uniform speed, after balancing, centrifuge at 500 g, 30 min, RT, slowly rise and fall;

[0101] 1.3) Extrac...

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Abstract

The invention relates to a co-culture method of a DC cell and a CIK cell and belongs to the technical field of cell culture. The co-culture method comprises the following steps: 1) extracting a single nucleus cell; 2) culturing the CIK cell; 3) culturing the DC cell; 4) carrying out mixed culturing on the DC cell and the CIK cell; and 5) carrying out multiplication culture. According to the co-culture method disclosed by the invention, after the DC cell is stimulated to be mature and co-cultured with the CIK cell, a factor GM-CSF is added according to the concentration and replenished after culture for 12h, so that the activity of the DC cell is kept, the survival time of the DC cell in a CIK culture system is prolonged, and the activity of the DC cell is kept for a long time.

Description

technical field [0001] The invention relates to a co-culture method of DC cells and CIK cells, which belongs to the technical field of cell culture. Background technique [0002] CIK biological immunotherapy, as the first choice for the treatment of tumors in the 21st century, is widely used in the treatment of various solid tumors. CIK biological immunotherapy has: 1) Proliferation, which can be expanded in a large amount in a short period of time; 2) CIK cells are derived from themselves without any side effects on the human body; 3) Broad tumor killing spectrum, with killing effect on most tumors; 4) It has high antitumor activity and is not affected by immunosuppressants such as CSA and FK506; 5) It can effectively resist the apoptosis mechanism caused by tumor cells. Dendritic cells, referred to as DC cells, are professional antigen-presenting cells with the most powerful functions in the body. Tumor cell immunity. [0003] DC cells are the initiators of the body's i...

Claims

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Application Information

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IPC IPC(8): C12N5/0784C12N5/0783
Inventor 闫敬武杜成德程思博孟艳芬
Owner 山东世博金都药业有限公司
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