Kit for separated culture of DC-CIK cells, and application thereof

A DC-CIK, isolation and culture technology, applied in the field of kits for the isolation and culture of DC-CIK cells, can solve the problems of low proportion of double-positive cells, fever, inflammation, and small number of patients, and achieve simple operation methods and good clinical practice Effect, the effect of fast proliferation

Inactive Publication Date: 2013-03-20
JIANGYIN CHI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestically, the lymphocyte separation method is mainly used. The purity and activity of the isolated DC and CIK cells are generally low, and there are more red blood cells, tumor infiltrating lymphocytes, platelets, etc. in them. When the cell fragments are returned to the human body, it is easy to cause patients to develop Adverse reactions such as fever and inflammation
[0006] (2) CIK cells proliferate slowly and the number is small
The proliferation rate of CIK cells cultivated by the existing domestic technology is slow, and it is difficult to reach the order of magnitude required for tumor treatment, and the tumor treatment effect is not good.
[0007] (3) The expression level of effective tumor suppressor factors in CIK cells is low
The cytotoxicity of CIK cells is positively correlated with the expression level of CD3+CD56+, while the proportion of CD3+CD56+ double-positive cells in CIK cells cultured by the existing domestic technology is low, and the double-positive expression rate is less than 10%

Method used

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  • Kit for separated culture of DC-CIK cells, and application thereof
  • Kit for separated culture of DC-CIK cells, and application thereof
  • Kit for separated culture of DC-CIK cells, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The composition of the DC-CIK cell isolation and culture kit in this embodiment is as follows:

[0049] (1) Lymphocyte separation fluid;

[0050] (2) Peripheral blood sample treatment solution: D-PBS buffer solution containing 1% bovine serum albumin by mass;

[0051] (3) CIK cell proliferation induction system: CD3, interleukin-1 and interleukin-2;

[0052] (4) DC induction proliferation system: GM-CSF, interleukin-4;

[0053] (5) Cell culture flask coating system: fibronectin, gamma interferon;

[0054] (6) Lymphocyte medium GT-T551.

[0055] Apply the above kit to the isolation and culture of DC-CIK cells, the specific steps are as follows:

[0056] (1) Separation of mononuclear cells from peripheral blood samples: draw 20mL of lymphocyte separation solution into a 50mL centrifuge tube, take 20mL of heparin-added blood bank concentrated white blood cells, and add 8mL of peripheral blood sample processing solution (containing mass percent D-PBS buffer solution w...

Embodiment 2

[0066] The composition of the DC-CIK cell isolation and culture kit in this embodiment is as follows:

[0067] (1) Lymphocyte separation medium

[0068] (2) Peripheral blood sample treatment solution: D-PBS buffer solution containing 2.5% bovine serum albumin by mass;

[0069] (3) CIK cell proliferation induction system: CD3, interleukin-1 and interleukin-2;

[0070] (4) DC induction proliferation system: GM-CSF, interleukin-4;

[0071] (5) Cell culture flask coating system: fibronectin, gamma interferon;

[0072] (6) Lymphocyte medium GT-T551.

[0073] Apply the above kit to the isolation and culture of DC-CIK cells, the specific steps are as follows:

[0074] (1) Separation of mononuclear cells from peripheral blood samples: draw 20mL of lymphocyte separation solution into a 50mL centrifuge tube, take 20mL of heparin-added blood bank concentrated white blood cells, and add 10mL of peripheral blood sample processing solution (containing mass percent D-PBS buffer solutio...

Embodiment 3

[0084] The composition of the DC-CIK cell isolation and culture kit in this embodiment is as follows:

[0085] (1) Lymphocyte separation medium

[0086] (2) Peripheral blood sample treatment solution: D-PBS buffer solution containing 3% bovine serum albumin by mass;

[0087] (3) CIK cell proliferation induction system: CD3, interleukin-1 and interleukin-2;

[0088] (4) DC induction proliferation system: GM-CSF, interleukin-4;

[0089] (5) Cell culture flask coating system: fibronectin, gamma interferon;

[0090](6) Lymphocyte medium GT-T551.

[0091] Apply the above kit to the isolation and culture of DC-CIK cells, the specific steps are as follows:

[0092] (1) Separation of mononuclear cells from peripheral blood samples: draw 20mL of lymphocyte separation solution into a 50mL centrifuge tube, take 20mL of heparin-added blood bank concentrated white blood cells, and add 15mL of peripheral blood sample processing solution (containing mass percent D-PBS buffer solution w...

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Abstract

The invention provides a kit for separated culture of DC-CIK cells, and an application thereof. The kit comprises a lymphocyte separation liquid, a peripheral blood sample treatment liquid, a CIK cell induced propagation system, a DC induced propagation system, a cell culture bottle coating system, a lymphocyte culture medium GT-T551 and a cell culture bag. The application of the kit in separating and culturing the DC-CIK cells comprises the following steps of separation of peripheral blood sample mononuclear cells, separation of the DC cells and CIK cells, induced propagation of the DC cells, the induced propagation of the CIK cells and co-culture of the DC cells and CIK cells. The lymphocytes obtained from the separation by the kit have very high purity and activity; the propagation rate of the CIK cells is fast; the proportion of CD3+CD56+ double positive cells is high; operation method is simple; and conditions are easy to control. The kit can be widely applied in the separated culture for the DC-CIK cells of human and mammals.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for isolating and cultivating DC-CIK cells and an application thereof. Background technique [0002] Biological therapy centered on tumor immunotherapy has become the fourth treatment mode after surgery, radiotherapy and chemotherapy. This treatment mode uses the patient's own immune cells to induce, stimulate or assist the body's own immune system, thereby improving and enhancing the patient's immune function, and the treatment method is safe and basically has no toxic side effects, so it has shown good application prospects in recent years. [0003] DC-CIK combined therapy is one of the most mature treatment options in tumor biological therapy. DC (dendritic cell) is the most powerful antigen-presenting cell discovered so far. It can efficiently and accurately identify tumor antigens and transmit the information to the human immune system. It plays an important r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/078
Inventor 齐来俊
Owner JIANGYIN CHI SCI
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