Method for enhancing reproductive capacity of CIK (Cytokine Induced Killer) cell and improving tumor cell killing capacity

A technology of cell proliferation and ability, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems of low number and activity of CIK cells, low tumor cell killing rate, poor therapeutic effect, etc., to improve cell Toxic effect, the effect of enhancing the growth ability

Inactive Publication Date: 2013-03-20
深圳市中美康士生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the number and activity of CIK cells obtained by the current method of cultivating CI

Method used

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  • Method for enhancing reproductive capacity of CIK (Cytokine Induced Killer) cell and improving tumor cell killing capacity
  • Method for enhancing reproductive capacity of CIK (Cytokine Induced Killer) cell and improving tumor cell killing capacity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Aseptically collect peripheral blood from healthy people and send it back to the cell preparation laboratory for experiments in a clean bench; separate mononuclear cells with lymphatic separation fluid, wash twice, resuspend cells with Q007 medium, and adjust the cell density at 3*10 6 / ml, and then follow the steps and methods below to culture CIK cells and control experiments.

[0024] The experiment was divided into a blank group, a phytohemagglutinin group and a positive control CD3McAb group. The design scheme is as follows:

[0025] The experiment was carried out in a 12-well plate, and the inoculum volume in each well was 1ml

[0026] Blank control group:

[0027] Day 0: IFN-γ (250IU / ml)

[0028] Day 1: IL-1 (2.5ng / ml)

[0029] Day 2: IL-2 (1000IU / ml)

[0030] Remaining days: IL-2 (1000IU / ml)

[0031] Positive control group:

[0032] Day 0: IFN-γ (250IU / ml)

[0033] Day 1: CD3McAb (50ng / ml) 20ul, IL-1 (2.5ng / ml)

[0034] Day 2: IL-2 (1000IU / ml)

[0035] ...

Embodiment 2

[0053] Aseptically collect peripheral blood from healthy people and send it back to the cell preparation laboratory for experiments in a clean bench; use lymph separation fluid to separate mononuclear cells, resuspend the cells with Q007 medium, adjust the cell density to 2*106 / ml, and then The CIK cell culture and experimental control were carried out according to the following steps and methods.

[0054] The experiment was divided into a blank group, a phytohemagglutinin group and a positive control CD3McAb group. The design scheme is as follows:

[0055] The experiment was carried out in a 12-well plate, and the inoculum volume in each well was 1ml

[0056] Blank control group:

[0057] Day 0: IFN-γ (250IU / ml)

[0058] Day 1: IL-1 (2.5ng / ml)

[0059] Day 2: IL-2 (1000IU / ml)

[0060] Remaining days: IL-2 (1000IU / ml)

[0061] Positive control group:

[0062] Day 0: IFN-γ (250IU / ml)

[0063] Day 1: CD3McAb (50ng / ml) 20ul, IL-1 (2.5ng / ml)

[0064] Day 2: IL-2 (1000IU / ml...

Embodiment 3

[0074] Aseptically collect peripheral blood from healthy people and send it back to the cell preparation laboratory for experiments in a clean bench; separate mononuclear cells with lymphatic separation fluid, wash twice, resuspend cells with Q007 medium, and adjust the cell density at 3*10 6 / ml, and then follow the steps and methods below to culture CIK cells and control experiments.

[0075] The experiment was divided into a blank group, a phytohemagglutinin group and a positive control CD3McAb group. The design scheme is as follows:

[0076] The experiment was carried out in a 12-well plate, and the inoculum volume in each well was 1ml

[0077] Blank control group:

[0078] Day 0: IFN-γ (250IU / ml)

[0079] Day 1: IL-1 (2.5ng / ml)

[0080] Day 2: IL-2 (1000IU / ml)

[0081] Remaining days: IL-2 (1000IU / ml)

[0082] Positive control group:

[0083] Day 0: IFN-γ (250IU / ml)

[0084] Day 1: CD3McAb (50ng / ml) 20ul, IL-1 (2.5ng / ml)

[0085] Day 2: IL-2 (1000IU / ml)

[0086] ...

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Abstract

The invention provides a method for enhancing the reproductive capacity of CIK (Cytokine Induced Killer) cells and improving tumor cell killing capacity. The method comprises the following steps of: sterilely collecting and separating the peripheral blood mononuclear cells of a healthy person; carrying out resuspension on the obtained peripheral blood mononuclear cells by using a Q007 culture medium; adding 24-26 ug/ml of PHA-L type phytohemagglutinin; and culturing for at least 15 days in the Q007 culture medium. The method provided by the invention has the beneficial effects that the reproductive capacity of the CIK cells and the cytotoxic effect are enhanced by adding 24-26 ug/ml of the PHA-L type phytohemagglutinin in a CIK cell culturing process, thereby enhancing the kill rate and the cure effect of the tumor cells.

Description

technical field [0001] The present invention relates to a method for culturing cytokine-induced killer cells (CIK cells). Background technique [0002] Tumor is a major disease that endangers human life and health, and it has been listed as the second cause of death in the population. According to the statistics of the World Health Organization, there are 9 million new cancers and 5 million deaths worldwide every year. The current treatment methods for tumors are mainly surgery, radiotherapy and chemotherapy. Among them, surgical treatment is only suitable for early-stage tumors, while radiotherapy and chemotherapy are more toxic and side effects. Therefore, developing a new generation of safe and effective anti-tumor biological therapy will have extremely important social and economic benefits. [0003] Stanford University in the United States first reported a type of killer cells induced by a variety of cytokines, namely, cytokine-induced killer cells (cytokine-induced ...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
Inventor 李晓祥沈政黄谋珍
Owner 深圳市中美康士生物科技有限公司
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