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Medium composition for culturing self-activated lymphocytes and method for culturing self-activated lymphocytes using same

A technology of self-excited lymphocytes and culture methods, which is applied in the field of self-excited lymphocyte culture medium composition and self-excited lymphocyte culture, can solve the problems of limited killing effect of cancer cells, and achieve simple implementation methods and low side effects. little effect

Inactive Publication Date: 2013-05-01
CELLS SCI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, when this method is applied to the treatment of tumors, although the cancer cells that remember the event can be effectively attacked and killed by a large number of activated T cells, the killing effect on various cancer cells that are not remembered is limited. limitation

Method used

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  • Medium composition for culturing self-activated lymphocytes and method for culturing self-activated lymphocytes using same
  • Medium composition for culturing self-activated lymphocytes and method for culturing self-activated lymphocytes using same
  • Medium composition for culturing self-activated lymphocytes and method for culturing self-activated lymphocytes using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1: Manufacture of self-excited lymphocytes

[0071] With 60cc of lymphocytes taken from the patient's peripheral blood 1×10 7 After the suspension was supported by 1ml of culture medium, it was mixed into 9ml of culture medium supplemented with IL-2, L-glutamine, and autologous plasma. In the presence of IL-12, IL-18, anti-CD3 antibody, and anti-CD16 antibody Under the condition of , cultured for 4 days, and carried out the first culture stage.

[0072] The culture medium used in the first culture stage here is based on 39ml of culture medium. As the nutritional ingredients required for immune cell culture, 200mM L-glutamine 400ul and 4ml of autologous plasma are added; as immune cell activation components, 18×10 6 IU / ml of IL-24ul, 100ug / ml of IL-124ul, 100ug / ml of IL-1820ul; as the final cultured immune cell relative ratio adjustment components, add 1mg / ml of anti-CD3 and anti-CD16 antibody 2.5 each ul culture medium.

[0073] Next, mix the culture mixtur...

Embodiment 2

[0077] Example 2: Observation of changes in phenotype and cell number before and after culture

[0078] image 3 and Figure 4 It is a schematic diagram of the phenotype changes of lymphocytes before and after culture. The H1 area is NK cells, the H4 area is T cells, the H2 area is NKT cells, and the H3 area is the distribution area of ​​other immune cells. Figure 5 It is a schematic diagram comparing the change of NK cell number obtained by the self-stimulated lymphocyte culture method of the present invention with the NK cell number change obtained by the self-stimulated lymphocyte culture method of the prior art.

[0079] The surface antigens of the activated lymphocytes cultured by the method of Example 1 were analyzed by flow cytometry. results, such as image 3 , Figure 4 As shown, before culturing as image 3 As shown, the most distributed surface antigen is in the H4 region, and after culture as Figure 4 As shown, the most surface antigen distribution is the...

Embodiment 3

[0082] Example 3: Analysis of cytotoxicity to various cancer cells

[0083] Figure 6 It is a schematic diagram showing the comparison of the cytotoxicity analysis results of activated lymphocytes cultured by the self-activated lymphocyte culture method of the present invention and the cytotoxicity analysis results of activated lymphocytes cultured by the self-activated lymphocyte culture method of the prior art.

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Abstract

Disclosed is a medium composition for culturing self-activated lymphocytes, which contains anti-CD3 antibody and anti-CD16 antibody in addition to interleukin 2 (IL-2), interleukin 12 (IL-12) and interleukin 18 (IL-18) in a medium, and thus can efficiently proliferate and activate NK cells, T cells and NKT cells and, at the same time, can significantly increase the ratio of NK cells in lymphocytes so as to provide immunocytes having excellent effects on the treatment of various kinds of malignant tumors, and a method for culturing self-activated lymphocytes using the medium composition.

Description

technical field [0001] The present invention relates to a medium composition for cultivating self-excited lymphocytes for treating malignant tumors and a method for culturing self-excited lymphocytes, in particular to a medium in which interleukin 2 (IL-2) and interleukin 12 (IL-12) and interleukin 18 (IL-18) are also added with CD3 and CD16 antibodies, which can effectively increase and activate NK (natural killer) cells, T cells and NKT cells, and significantly increase the NK cells in lymphocytes. The composition of the culture medium for self-stimulating lymphocytes capable of cultivating immune cells with excellent curative effect on treating various malignant tumors, and the self-stimulating lymphocyte culture method using the same. Background technique [0002] In recent anticancer treatments, adoptive immunotherapy (adoptive immunotherapy) that can replace conventional surgery, radiation therapy, and chemotherapy has attracted attention. [0003] Adoptive immunother...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078C12N5/02C07K14/54C07K16/28
CPCA61K2039/57C12N2501/515C12N2501/2302A61K39/0011C12N2501/2312C12N5/0636C12N1/38A61K2039/5158C12N2501/2318C07K16/283C12N2501/599C07K16/2809C12N5/0638C12N5/0646A61K39/4611A61K39/4644A61K39/4613
Inventor 洪鲜旼白咏锡
Owner CELLS SCI CORP
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