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Immune cell culture medium and culture method, as well as application of ginsenoside

A technology of immune cells and ginsenosides, applied in the application of ginsenosides, the field of immune cell culture medium, can solve the problems of poor immune cell activity and proliferation ability, etc.

Inactive Publication Date: 2015-11-25
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] It can be seen that the activity and proliferation ability of immune cells cultured in the existing cell culture medium are poor

Method used

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  • Immune cell culture medium and culture method, as well as application of ginsenoside
  • Immune cell culture medium and culture method, as well as application of ginsenoside
  • Immune cell culture medium and culture method, as well as application of ginsenoside

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preparation example Construction

[0033] The immune cell culture medium provided by the invention includes ginsenoside, and the mass content of the ginsenoside in the serum-free basal medium is 4 mg / mL-10 mg / mL. In a specific embodiment of the present invention, the mass content of the ginsenosides in the serum-free basal medium can be 4 mg / mL, or 6 mg / mL, or 10 mg / mL; the present invention has no special restrictions on ginsenosides. Restrictions, the use of ginsenosides well known to those skilled in the art can be used, such as commercially available products can be used, or the preparation technical scheme for preparing ginsenosides well known to those skilled in the art can be used to prepare by itself. In the present invention, the preparation method of the ginsenoside preferably comprises the following steps:

[0034] extracting the coarse powder of ginseng stems and leaves, and filtering to obtain a primary filtrate;

[0035] The filtrate is removed from impurities, and then filtered to obtain a secon...

Embodiment 1

[0075] Take 40mL of peripheral blood, and use Ficoll separation method to obtain PBMC (mononuclear cells) 4×10 7 , divided into 10 groups, each group 4×10 6 Cells were inoculated in X-VIVO15 serum-free medium (purchased from LONZA Company), and ginsenosides were added thereto so that the concentrations of ginsenosides were 1 mg / mL, 2 mg / mL, 3 mg / mL, and 4 mg / mL, respectively. , 5mg / mL, 6mg / mL, 7mg / mL, 8mg / mL, 9mg / mL, 10mg / mL;

[0076] The inoculation density was 1×10 6 cells / mL; on the first day after inoculation, the cytokine INF-γ was added to a final concentration of 30 ng / mL; 80ng / mL;

[0077] Rehydration was performed every 3 days, and cytokine IL-2 was added according to the total volume after rehydration to a final concentration of 300 U / mL; after 14 days of culture, proliferated CIK cells were obtained.

[0078] The present invention counts the CIK cells after proliferation, and draws the curve of ginsenoside concentration and CIK cell proliferation multiple, such ...

Embodiment 2

[0080] Take 40mL of peripheral blood, and use Ficoll separation method to obtain PBMC (mononuclear cells) 4×10 7 , divided into 10 groups, each group 4×10 6 cells;

[0081] Groups 1 and 2 were used as control groups 1 and 2: groups 1 and 2 were induced to culture CIK cells using the serum-free medium in Comparative Example 1;

[0082] Groups 3 and 4 were used as control groups 3 and 4: Groups 3 and 4 used X-VIVO15 serum-free medium produced by LONZA Company in the United States to induce and culture CIK cells;

[0083] The 5th to 10th groups were used as the experimental groups of the present invention, wherein, the 5th and 6th groups were inoculated in X-VIVO15 serum-free medium, induced and cultured CIK cells, and the content of ginsenoside was 4 mg / mL;

[0084] Groups 7 and 8 were inoculated in X-VIVO15 serum-free medium to induce and culture CIK cells with a ginsenoside content of 10 mg / mL;

[0085] Groups 9 and 10 were inoculated in X-VIVO15 serum-free medium to induce a...

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Abstract

The invention provides an immune cell culture medium and an immune cell culture method, as well as an application of ginsenoside. The immune cell culture medium comprises serum-free basal culture medium, ginsenoside and cytokines, wherein the weight content of ginsenoside in the serum-free basal culture medium is 4mg / mL-10mg / mL. The immune cell culture medium provided by invention can enhance the activity and the multiplication capacity of the immune cell. The experimental result shows that after CIK cells are cultured by the immune cell culture medium by 14 days, the proliferation rate is 650-750 times, the survival rate is 97.78-98.73%, the ratio of the cells with the CD3+CD56+ effects is 37.4%-38.2%; after NKT cells are cultured by the immune cell culture medium by 14 days, the proliferation rate is 480-550 times, the survival rate is 96.73-98.78%, and the ratio of the cells with the summation effects of CD3+CD56+ and CD161+ is 42.6%-45.2%.

Description

technical field [0001] The invention belongs to the technical field of immune cell culture, and in particular relates to an immune cell culture medium, a method for culturing immune cells and the application of ginsenoside. Background technique [0002] Immune cells refer to cells that participate in or are related to immune responses, and can be divided into various types, including lymphocytes, dendritic cells, monocytes / macrophages, granulocytes, mast cells, etc. In the human body, various immune cells play important roles. [0003] Immune cell therapy is to collect the body's own immune cells, culture them in vitro, increase their number by thousands of times, and enhance the targeted killing function, and then infuse them back into the human body to kill pathogens and cancer cells in blood and tissues. Cells and mutated cells can break immune tolerance, activate and enhance the immune ability of the body, and take into account the dual effects of treatment and health c...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12N5/0783C12N5/0784C12N5/0786C12N5/0787
Inventor 陈海佳王一飞葛啸虎应杰万桦
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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