Application of lipid compound combination in T cell culture

A cell culture and compound technology, applied in cell culture active agents, animal cells, tissue culture, etc., can solve the problem of not considering the proportion of CD4 and CD8 subgroups, and achieve no animal-derived components, clear chemical components, high clinical The effect of applying value

Pending Publication Date: 2021-06-22
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most prior art T cell culture methods do not consider the ratio of CD4 and CD8 subsets in the T cell population at the end of culture, or tend to maintain the ratio of CD4 and CD8 subsets in the initial T cell population

Method used

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  • Application of lipid compound combination in T cell culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Expansion of different proportions of CD4-positive and CD8-positive T cells in human peripheral blood mononuclear cells (PBMC) (1)

[0044] In this example, PBMCs were freshly isolated from human whole blood by density centrifugation for activation and expansion of T cells. Transfer the separated PBMC cell suspension to the prepared 6-well culture dish, seeding density 1x10 6 / ml, use X-Vivo-15 (Lonza) medium 2mL / well added with lipid compound combination solution, and add CD3 antibody at a final concentration of 10μg / mL, CD28 antibody at a final concentration of 5μg / mL, and a final concentration of 100IU / mL IL2, and placed in a 37°C, 5% carbon dioxide incubator to continue culturing.

[0045]Wherein, the solute of lipid compound combination solution is following 8 kinds of lipids: myristic acid, arachidonic acid, cholesterol, linoleic acid, oleic acid, palmitic acid, hard fatty acid, linolenic acid; Its concentration is 0.00125g / L.

[0046] Four conditio...

Embodiment 2

[0048] Example 2 Expansion of different proportions of CD4-positive and CD8-positive T cells in human peripheral blood mononuclear cells (PBMC) (2)

[0049] In this example, PBMCs were freshly isolated from human whole blood by density centrifugation for activation and expansion of T cells. Transfer the separated PBMC cell suspension to the prepared 6-well culture dish, seeding density 1x10 6 / ml, use OpTmizer (ThermoFisher) medium 2mL / well added with lipid compound combination solution, and add CD3 antibody at a final concentration of 10 μg / mL, CD28 antibody at a final concentration of 5 μg / mL, and IL2 at a final concentration of 100 IU / mL , and placed in a 37°C, 5% carbon dioxide incubator to continue culturing.

[0050] Wherein, the solute of the lipid compound combination solution is the following two kinds of lipids: myristic acid and arachidonic acid; the concentrations thereof are both 0.00125 g / L.

[0051] Four conditions were set up in the experiment, and the additi...

Embodiment 3

[0053] Example 3 Expansion of different proportions of CD4-positive and CD8-positive T cells in human peripheral blood mononuclear cells (PBMC) (3)

[0054] In this example, PBMCs were freshly isolated from human whole blood by density centrifugation for activation and expansion of T cells. Transfer the separated PBMC cell suspension to the prepared 6-well culture dish, seeding density 1x10 6 / ml, use X-Vivo-15 (Lonza) medium 2mL / well added with lipid compound combination solution, and add CD3 antibody at a final concentration of 10μg / mL, CD28 antibody at a final concentration of 5μg / mL, and a final concentration of 100IU / mL IL2, and placed in a 37°C, 5% carbon dioxide incubator to continue culturing.

[0055] Wherein, the solute of the lipid compound combination solution is the following four kinds of lipids: oleic acid, palmitic acid, stearic fatty acid, and linolenic acid; the concentrations thereof are all 0.0001 g / L.

[0056] There are 9 conditions in the experiment, a...

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Abstract

The invention relates to an application of a lipid compound combination in T cell culture. The lipid compound combination comprises any two or more lipid compounds of alpha-tocopheryl acetic acid, myristic acid, arachidonic acid, cholesterol, linoleic acid, oleic acid, palmitic acid, stearic acid, linolenic acid and palmitoleic acid. The lipid compound composition is added into a T cell culture medium according to different proportions, the proportion of CD4 and CD8 subgroups in a T cell population at the amplification end point can be adjusted at will, the requirements of scientific research or clinical application are met, and the lipid compound composition is definite in chemical component, the chemical component are a common nutrient substance in the cell culture process, safe and harmless, and free of human-derived and animal-derived components, and the clinical application requirements are met. The method provided by the invention has very high clinical application value.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to the application of a lipid compound combination in T cell culture. Background technique [0002] T cells are one of the main components of lymphocytes. They have a variety of biological functions, such as directly killing target cells, assisting other lymphocytes to function, responding to specific antigens or mitogens, and producing cytokines. One of the body's main immune cells that fight off infection and prevent tumors from forming. Enrichment, activation, and expansion of such immune cells in vitro can be used for reinfusion to treat a variety of diseases, including malignant tumors, infections, and autoimmune diseases. [0003] T cells can be divided into CD4 positive T cells and CD8 positive T cells according to the difference of surface molecular markers. Among them, CD4-positive T cells account for about 65%-70% of the total number of T cells, and CD8-positive T c...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2500/36C12N2500/38C12N2500/30C12N2500/90C12N2501/2302C12N2501/51C12N2501/515C12N2500/02
Inventor 陈涛涛陈旭陈刚檀灯华林家会辛丽丽
Owner 苏州依科赛生物科技股份有限公司
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