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Improved RPMI1640 culture medium and application thereof

A technology of RPMI1640 and culture medium, which is applied in the field of improved RPMI1640 medium, can solve the problems of affecting T cell killing ability and T cell exhaustion, and achieve the effect of improving killing function, improving tumor killing ability and avoiding excessive activation

Active Publication Date: 2020-02-07
WUHAN UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, during the process of CAR-T cell expansion in vitro, T cells will be exhausted, which will affect the killing ability of T cells

Method used

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  • Improved RPMI1640 culture medium and application thereof
  • Improved RPMI1640 culture medium and application thereof
  • Improved RPMI1640 culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The medium components of embodiment 1-embodiment 3 and comparative example are shown in the table below:

[0030]

[0031] The culture medium of Example 1-Example 3 is an improved RPMI1640 medium, represented by MTM1-MTM3; the comparative example is RPMI1640 medium.

[0032] The preparation method of each medium in the above-mentioned Examples 1-Example 3 and the comparative example includes: mixing the components of the above-mentioned medium with high-purity sterilized water in proportion, shaking and mixing at 4°C for 1 hour, and then passing through a 0.22 μm Sterilize by membrane filtration.

[0033] The following experiments using the culture medium of Examples 1-3 and comparative examples were repeated in three cases of T cells from healthy adults.

[0034] Isolation of PBMCs and T cells: Peripheral blood was collected from healthy adult donors, PBMCs were separated by Ficoll, and T cells were separated by anti-CD3 magnetic beads.

[0035] The preparation me...

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PUM

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Abstract

The invention provides an improved RPMI1640 culture medium. The concentrations of sodium chloride and potassium chloride in the RPMI1640 culture medium are changed. Compared with the RPMI1640 culturemedium, when the culture medium is used for culturing the CAR-T cells, the normal proliferation of the CAR-T cells can be kept, meanwhile, the exhaustion of the CAR-T cells can be reduced, the anti-tumor activity of the CAR-T cells can be enhanced, and the culture medium has a good application prospect.

Description

technical field [0001] The invention relates to the field of biomedical cell therapy, in particular to an improved RPMI1640 culture medium and its application. Background technique [0002] According to the statistics of the Global Cancer Report, blood cancers such as leukemia, lymphoma, and multiple myeloma seriously threaten human life and health, ranking eighth among the top ten common malignant tumors, and the incidence rate is increasing year by year. In recent years, immunotherapy has brought new hope for the treatment of human hematological tumors. Among them, genetic engineering technology is used to transform T cells so that they can express antibodies that can recognize tumor antigens, target and recognize tumor cells, activate T cells and kill tumors Cell, that is, chimeric antigen receptor T cell (Chemeric Antigen Receptor T cell, CAR-T) immunotherapy, is the most promising tumor immunotherapy and research hotspot in recent years. At present, CAR-T cell immunoth...

Claims

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Application Information

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IPC IPC(8): C12N5/10
CPCC12N5/0636C12N2500/12C12N2500/14C12N2500/16C12N2500/30C12N2500/32C12N2500/34C12N2500/35C12N2500/38C12N2500/84C12N2510/00
Inventor 包乐媛李会廖兴华张同存
Owner WUHAN UNIV OF SCI & TECH
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