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Culture medium, application thereof and culture method

A culture method and culture medium technology, applied in the culture medium and its application and culture field, can solve the problems of reduced expression of chimeric antigen receptors, limited culture days, and inability of T cells to proliferate in large quantities, and achieve the effect of increasing the multiplication factor.

Active Publication Date: 2015-12-16
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] 1. The transfected T cells cannot proliferate in large quantities because of the risk of gene loss, usually CD4 + and CD8 + The cell proliferation factor is about 50 times;
[0009] 2. The number of cultivation days is limited, usually only about 7 days;
[0010] 3. The expression of chimeric antigen receptors in cells will decrease after culture. Taking T cells transfected with CD19CAR as an example, even if the transfection efficiency reaches more than 90%, the proportion of cells expressing CD19CAR will drop to 60% after 7 days of culture. %about

Method used

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  • Culture medium, application thereof and culture method
  • Culture medium, application thereof and culture method
  • Culture medium, application thereof and culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The preparation of embodiment 1 culture medium

[0059] The medium formula is shown in Table 1:

[0060] Table 1 medium formula

[0061] ingredient name

[0062] Culture medium preparation method: Mix the components of the above medium in proportion, shake and mix at 4°C for 1 hour, then filter and sterilize through a 0.22 μm filter membrane for use.

Embodiment 2

[0063] The preparation of embodiment 2 culture medium

[0064] The medium formula is shown in Table 2:

[0065] Table 2 medium formula

[0066] ingredient name

[0067] Vitamin C

[0068] Culture medium preparation method: Mix the components of the above medium in proportion, shake and mix at 4°C for 1 hour, then filter and sterilize through a 0.22 μm filter membrane for use.

Embodiment 3

[0069] The preparation of embodiment 3 medium

[0070] The medium formula is shown in Table 3:

[0071] Table 3 medium formula

[0072] ingredient name

[0073] Culture medium preparation method: Mix the components of the above medium in proportion, shake and mix at 4°C for 1 hour, then filter and sterilize through a 0.22 μm filter membrane for use.

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Abstract

The invention relates to the technical field of cell culture, in particular to a culture medium, application thereof and a culture method. The culture medium comprises transferrin, recombinant human insulin, progesterone, fetal calf serum, stem cell growth factors, vitamin C, interleukin 2, interleukin 4, zinc sulfate and a DMEM / F12 culture medium. When the culture medium is used for culturing CD19-CAR-T cells, the cell proliferation multiple can be improved, the number of cell culture days is increased, and the CD19CAR and other surface markers can be kept expressed.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a culture medium and its application and culture method. Background technique [0002] Mantle cell lymphoma (Mantle cell lymphoma) is a kind of non-Hodgkin's lymphoma (NHL), and about 70% of patients are already in stage III or stage Ⅳ disease at the time of diagnosis. The clinical classic CHOP (cyclophosphamide + doxorubicin + vincristine + prednisone) regimen is not satisfactory for the treatment of mantle lymphoma, only a few patients achieve complete remission, so new treatment strategies are urgently needed to Improve efficacy. [0003] With the development of tumor immunology theory and technology, adoptive cell immunotherapy has made considerable progress in recent years, among which the achievement of tumor-targeted immunotherapy represented by chimeric antigen receptor (chimeric antigen receptor, CAR) modified T cells is particularly important. Outstanding, in vit...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
Inventor 陈海佳王一飞葛啸虎万桦
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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