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A medium composition for cultivating self activated lymphocyte

A technology for cell culture and lymphocytes, which is applied in the direction of cell culture active agents, animal cells, tissue culture, etc., can solve the problems of attacking and restricting cancer cells, and achieve the effects of improving prognosis, easy administration, and less side effects

Inactive Publication Date: 2009-12-16
NKBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, this kind of NK cells with a high ability to kill cancer cells only accounts for 5% to 15% of normal peripheral blood lymphocytes, especially in the case of cancer patients, this ratio drops to less than 1%. The additional amplification process of immunotherapy makes it impossible to effectively attack cancer cells
[0008] However, the culture medium composition used in the above-mentioned method for activating CD4 T cells has the following technical problems; because the culture medium composition can only selectively activate T cells involved in acquiring immunity among immune cells, when using this method When treating tumors, although a large number of activated T cells can effectively attack or kill cancer cells that have been memorized, it is limited in the treatment of malignant tumors by attacking various cancer cells that have not been memorized

Method used

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  • A medium composition for cultivating self activated lymphocyte
  • A medium composition for cultivating self activated lymphocyte
  • A medium composition for cultivating self activated lymphocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Cell Proliferation and Culture of Lymphocytes

[0034] figure 1 It is a schematic diagram showing changes in the number of immune cells after cultured with the medium composition of the present invention.

[0035] Each lymphocyte isolated from the peripheral blood 60cc of a normal person, a solid tumor patient and a leukemia patient is used as an experimental group, and each lymphocyte is cultured in 39ml of the first culture medium composition for 4 days , and then mixed the first medium composition including the cultured cells into 67 ml of the second medium composition, and continued culturing for 2 days. In addition, in order to induce massive proliferation of cells, 7 ml of self-derived plasma was added to the supplemented cells and the second medium composition, and injected into 1 L of IL-2 containing 10 to 20 IU (International Unit) / ml. The cells were cultured for 8 days in a gas-permeable culture bag with a medium for cell culture. Here, each cu...

Embodiment 2

[0037] Embodiment 2: Observation of phenotype before and after culture

[0038] figure 2 It is a schematic diagram of the phenotype changes of activated lymphocytes cultured according to the medium composition of the present invention, the H1 area represents NK cells, the H4 area represents T cells, the H2 area represents NKT cells, and the H3 area represents the distribution of other immune cells.

[0039] Isolate lymphocytes from the peripheral blood of 60cc normal people, after the culture process of embodiment 1, utilize flow cytometer (floweytometry) to analyze the activated lymphocyte surface antigen of cultivating, use figure 2 To illustrate the results, as shown in Figure (a), the distribution of surface antigens before culture is mainly in the H4 region, and as shown in Figure (b), the distribution of surface antigens after culture is mainly in the H1 region, and the actual calculation of CD3 positive (Positive ) T cells and CD16+CD56 positive (positive) NK cells...

Embodiment 3

[0041] Example 3: Toxicity (cytotoxicity) analysis of various cancer cells

[0042] Lymphocytes were isolated from the peripheral blood of 60cc lymphosarcoma (lymphosarcoma) patients, and activated lymphocytes were cultivated through the culture process of the aforementioned embodiment 1, and cancer cells as target cells (target cells) were mixed with effector cells ( After activating lymphocytes of effector cell, after 6 hours, it was reacted with lactate dehydrogenase (lactatedehydrogenase; LDH), and the cytotoxicity was analyzed by enzyme-linked immunosorbent assay (Elisa), and the results were as follows: image 3 The activated self The cancer cell killing ability of activated lymphocytes was significantly improved compared with peripheral blood mononuclear cells (PBMCs).

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Abstract

Provided is a medium composition for culturing self- activated lymphocytes applicable to the treatment of malignant tumors, to which anti-CD3, anti-CD16 and anti-CD56 antibodies are added along with interleukin2 (IL-2) to evenly activate natural killer (NK) cells, T cells and natural killer T (NKT) cells, and thus the medium composition can be used to culture im- munocytes that can effectively treat various kinds of malignant tumors. The medium composition includes a cell culture medium and additives added to the cell culture medium, wherein the additives include interleukin2 (IL-2), anti-CD3 antibodies, anti-CD16 antibodies, and anti-CD56 antibodies.

Description

technical field [0001] The invention relates to a culture medium composition for cultivating autoactivated lymphocytes for treating malignant tumors. More specifically, the present invention relates to a culture medium composition, which contains anti-CD3, anti-CD16 and anti-CD56 antibodies in addition to interleukin 2 (IL-2), and can uniformly proliferate and activate NK cells, T cells, and NKT cells, so that immune cells that have significant functions in the treatment of a variety of malignant tumors can be cultivated. Background technique [0002] Recently, in cancer treatment, a new therapy called adaptive immunotherapy (adaptive immunotherapy), which can replace conventional surgery, radiation therapy, and chemotherapy, is attracting attention. [0003] Adoptive immunotherapy is a kind of immune system that obtains natural killer cells (natural killer cells; NK cells), dendritic cells (De), B cells, T cells, etc. from the patient's blood. Cells use a variety of stimu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/0783
CPCC12N2501/515C12N2501/23A61K39/0011C12N5/0646C12N5/0636A61K2039/5158C12N2501/599C12N5/0634A61K2239/48A61K39/461A61K39/464499
Inventor 李东旭洪藓旼崔炳仁
Owner NKBIO
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