Anti-drug-inhibition chimeric antigen receptor T cell culture medium composition and application thereof

A chimeric antigen receptor and culture medium technology, applied in the field of cell culture medium, can solve the problems of reduced CAR-T cell persistence, CAR loss, limited culture time, etc., and achieve the effect of increasing the killing activity and the expansion multiple.

Pending Publication Date: 2021-02-12
苏州米苏生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, serum-containing medium (such as IL-2+RPMI1640+FBS medium) is mainly used to cultivate CAR-T cells, but it has many shortcomings, such as: CAR-T cells cannot proliferate in large quantities after transfection, and CAR-T cells are prone to loss. This phenomenon leads to the gradual loss of CAR molecules on the surface of T cells after long-term culture; the limited culture time makes CAR-T cells unable to reach the number of reinfusion; and, with the progress of cell culture, T cells gradually change from memory T cells to effector cells. T cell transformation, which results in less persistence of CAR-T cells after entering the body

Method used

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  • Anti-drug-inhibition chimeric antigen receptor T cell culture medium composition and application thereof
  • Anti-drug-inhibition chimeric antigen receptor T cell culture medium composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 A CAR-T cell culture medium, consisting of RPMI1640 medium (commercially available) as a basic component and additional components. Specifically, the composition includes 500IU / ml IL-2, 600IU / ml IL-7, 100IU / ml IL-15, 5wt% B27, 4wt% autologous plasma, 25μg / mL modified molybdenum disulfide nanosheets, and the balance RPMI1640 medium. Wherein, the modified molybdenum disulfide nanosheets include molybdenum disulfide nanosheets and aniline dimers with a mass ratio of 10:1, and the aniline dimers are physically bound to the surface of the molybdenum disulfide nanosheets. The diameter of the molybdenum disulfide nanosheet is more than 1 μm, and the thickness is 1-2 nm. The modified molybdenum disulfide nanosheets can be prepared by the following method, that is: at room temperature, the molybdenum disulfide nanosheets and aniline dimer are dispersed in DMSO, and continue to stir for 1-2h, then with 10000-12000rpm Centrifuge at high speed, wash the precipitate with...

Embodiment 2

[0028] Example 2 A CAR-T cell culture medium, consisting of RPMI1640 medium (commercially available) as a basic component and additional components. Specifically, the composition includes 800IU / ml IL-2, 300IU / ml IL-7, 50IU / ml IL-15, 1wt% B27, 2wt% autologous plasma, 10μg / mL modified molybdenum disulfide nanosheets, and the balance RPMI1640 medium. Wherein, the modified molybdenum disulfide nanosheets include molybdenum disulfide nanosheets and aniline dimers with a mass ratio of 10:3, and the aniline dimers are physically bound to the surface of the molybdenum disulfide nanosheets. The diameter of the molybdenum disulfide nanosheet is more than 1 μm, and the thickness is 1-2 nm. The preparation method of the modified molybdenum disulfide nanosheets is the same as in Example 1.

Embodiment 3

[0029] Example 3 A CAR-T cell culture medium, consisting of RPMI1640 medium (commercially available) as a basic component and additional components. Specifically, the composition includes 600IU / ml IL-2, 500IU / ml IL-7, 80IU / ml IL-15, 3wt% B27, 3wt% autologous plasma, 20μg / mL modified molybdenum disulfide nanosheets, and the balance RPMI1640 medium. Wherein, the modified molybdenum disulfide nanosheets include molybdenum disulfide nanosheets and aniline dimers with a mass ratio of 5:1, and the aniline dimers are physically bound to the surface of the molybdenum disulfide nanosheets. The diameter of the molybdenum disulfide nanosheet is more than 1 μm, and the thickness is 1-2 nm. The preparation method of the modified molybdenum disulfide nanosheets is the same as in Example 1.

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Abstract

The invention discloses a chimeric antigen receptor T cell culture medium composition for resisting drug inhibition and application thereof. The composition comprises 500-800 IU/ml of IL-2, 300-600 IU/ml of IL-7, 50-100 IU/ml of IL-15, 1-5wt% of B27, 2-4wt% of autologous plasma, 10-25 [mu]g/mL of a modified molybdenum disulfide nanosheet, and the balance of an RPMI1640 culture medium. Compared with the prior art, when the CD19-CAR-T cells cultured by the culture medium disclosed by the invention are combined with chemical drugs to treat tumors, the CD19-CAR-T cells are less inhibited by the chemical drugs, and the survival rate, the killing activity and the amplification multiple of the cells are remarkably improved.

Description

technical field [0001] The invention relates to a cell culture medium, in particular to a drug-inhibited chimeric antigen receptor T cell culture medium composition and application thereof. Background technique [0002] Chimeric antigen receptor (CAR) T cell technology is a cell therapy technology that has developed very rapidly in recent years. Through genetic modification technology, the targeting, killing activity and persistence of effector T cells are higher than those of routinely used immune cells, and can overcome the local immunosuppressive microenvironment of the tumor and break the host immune tolerance state. CAR is composed of three parts: extracellular region, transmembrane region and intracellular region, which is essentially a membrane protein formed by series of different protein functional domains. [0003] At present, serum-containing medium (such as IL-2+RPMI1640+FBS medium) is mainly used to cultivate CAR-T cells, but it has many shortcomings, such as: ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10A61K35/17A61P35/00
CPCC12N5/0636A61K35/17A61P35/00C12N2510/00C12N2501/2302C12N2501/2307C12N2501/2315C12N2500/05
Inventor 吴建勇易小萍滕小锘张大鹤
Owner 苏州米苏生物技术有限公司
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