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Method for preparing CIK cell with killing effect on tumor cell

A tumor cell and cell technology, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems of high treatment costs, reduce the chance of cell contamination, and be easy to operate

Inactive Publication Date: 2013-07-03
UNION STEMCELL & GENE ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for patients, radiotherapy and chemotherapy are tantamount to drinking poison to quench their thirst. Patients not only need to pay high treatment costs but also endure great pain caused by side effects. What is even more sad is that this painful process only delays death.

Method used

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  • Method for preparing CIK cell with killing effect on tumor cell
  • Method for preparing CIK cell with killing effect on tumor cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Isolation of PBMCs:

[0058] 1) 50ml of fresh blood sample (peripheral blood or umbilical cord blood) and phosphate buffered saline (PBS) were all warmed to 20°C and mixed uniformly in a ratio of 1:1.

[0059] 2) Take three new 50ml centrifuge tubes, and add 15ml of Ficoll mononuclear cell separation solution (density 1.084) warmed to 20°C into each centrifuge tube.

[0060] 3) Use a disposable pipette to gently add 30ml of uniformly mixed blood dilution solution to the upper layer of Ficoll mononuclear cell separation solution and ensure that the two layers of liquid are clearly separated.

[0061] 4) Gently put the balanced centrifuge tube into the centrifuge and centrifuge at 250g for 40min.

[0062] 5) Gently take out the centrifuge tube, use a new pipette to first suck out the upper layer of serum to a new centrifuge tube for later use, then absorb the mononuclear cell buffy coat layer and 5ml of liquid above and below the buffy coat layer into another new centrif...

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Abstract

The invention discloses a method for preparing CIK cells with a killing effect on tumor cells. The CIK cells are obtained through induced culture of 17-20 days by using a ficoll mononuclear cell separation medium with a density of 1.084 in the PBS system to separate mononuclear cells in peripheral blood or umbilical cord blood. According to the present invention, mononuclear cells are separated and obtained efficiently by using the ficoll density gradient centrifugation method, and a sufficient amount of CIK is obtained by using cell culture bags and a CIK cell culture system to meet the needs of clinical treatment. The Takara lymphocyte culture medium and the autologous serum and cytokine co-culture technique are used in the method, thereby preventing application of fetal bovine serum, reducing contamination risks of exogenous pyrogen and sensitinogen, and while maintaining the advantage of CIK cell efficient proliferation. The cell culture bag technology is used to reduce risk of cell contamination, and is suitable for clinical therapeutic application.

Description

technical field [0001] The invention relates to a method for isolating mononuclear cells (PBMC) from human peripheral blood or umbilical cord blood, and obtaining CIK cells with tumor cell killing effect in the simplest way within 17-20 days. Background technique [0002] Tumor has always been almost synonymous with death. Although a lot of work has been done in the field of medicine and biology around the occurrence, mechanism and treatment of tumors, the most important treatment methods so far are radiotherapy and chemotherapy. However, for patients, radiotherapy and chemotherapy are tantamount to drinking poison to quench their thirst. Patients not only need to pay high treatment costs but also endure great pain caused by side effects. What is even more sad is that this painful process only delays death. . Therefore, relevant experts and scholars have increasingly realized that another route should be found to conquer tumors. [0003] Cell therapy has not only received...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
Inventor 陈晓波韩洪起韩俊领黄家学武文杰侯士芳张宇光
Owner UNION STEMCELL & GENE ENG
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