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Serum-free culture medium and culture method of CAR-T cells

A serum-free medium and culture method technology, applied in cell culture active agents, animal cells, culture process and other directions, can solve the problems affecting the cost of treatment of patients, and achieve the effect of improving cell survival rate, easy implementation and reducing culture cost.

Pending Publication Date: 2021-12-21
曾献芳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is necessary to culture and expand a large number of CAR-T cells in vitro. Generally, a patient needs hundreds of millions or even billions of CAR-T cells. greatly affect the cost of treatment for patients

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A Car-T cell-free-serum-free medium, which consists of X-Vivo 15 medium and the following components added to the medium: Ni Miao, 2.8 ng / ml, eprotcharin 8.2 ng / ml, ferulic acid ester 9.4 ng / mL, IL-2 150IU / mL, IL-15400 IU / mL, vitamin E 25 μg / ml, L-glutamine 15 μg / ml.

[0022] A CAR-T cell culture method, which encases the obtained CAR-T cells and the same amount of CD3 / CD28 magnetic beads, CAR-T cells concentration in medium in the medium 3 × 10 6 / Ml, at 37 ° C, 5% CO 2 The culture tank was cultured for 10 days, and a medium was supplemented once every 1 day during the culture.

Embodiment 2

[0024] A car-T cell-free-serum-free medium, which consists of X-Vivo 15 medium and the following ingredient added to the medium: Ni Miao, 2.8 ng / ml, Epiracein 6.8 ng / ml, ferulicate 8.5 ng / ml, IL-2 100 IU / mL, IL-15300 IU / mL, vitamin E 20 μg / mL, L-glutamine 10 μg / ml.

[0025] A CAR-T cell culture method, which encases the obtained CAR-T cells and the same amount of CD3 / CD28 magnetic beads, CAR-T cells concentration in medium in the medium 1 × 10 6 / Ml, at 37 ° C, 5% CO 2 10 days were cultured in the incubator, and a medium was supplemented every 2 days during the culture.

Embodiment 3

[0027] A car-T cell-free-serum-free medium, which consists of x-Vivo 15 medium and the following ingredient added to the medium: nigerin 3.5 ng / ml, eprotcharin 9.6 ng / ml, ferulicate 10 ng / ml, IL-2 200 IU / mL, IL-15500 IU / mL, vitamin E 30 μg / ml, L-glutamine 18 μg / ml.

[0028] A CAR-T cell culture method, which disseptes the obtained CAR-T cells and the same amount of CD3 / CD28 magnetic beads were added to the medium of T75 culture flasks, and CAR-T cells concentrated in the medium to 5 × 10 6 / Ml, at 37 ° C, 5% CO 2 The culture tank was cultured for 10 days, and a medium was supplemented once every 1 day during the culture.

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PUM

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Abstract

The invention discloses a serum-free culture medium of CAR-T cells. The serum-free culture medium consists of a basic culture medium and the following components added into the culture medium: nicergoline, icariin, ferulate, IL-2, IL-15, vitamin E and L-glutamine. The nicergoline and the icariin are added into the serum-free culture medium of the CAR-T cells in the invention, so that the proliferation activity of the CAR-T cells can be effectively improved; and a large number of CAR-T cells can be harvested within a short time. The ferulate is also added, so that the number of living cells in the harvested cells is easily increased; and the cell survival rate is increased. The components, the L-2, the IL-15, the vitamin E and the L-glutamine jointly act; the proliferation activity of the CAR-T cells is improved; the used components are safe; blood components such as plasma are not added; the culture cost of the CAR-T cells is reduced; and the burden of a patient is relieved. The invention further provides a culture method of the CAR-T cells; the culture method is simple and convenient to operate and easy to implement; and popularization and application of the CAR-T cells in the field of tumour immunotherapy are facilitated.

Description

Technical field [0001] The present invention relates to the field of immunocytes, and in particular, to a CAR-T cell-free medium serum-free medium and a culture method thereof. Background technique [0002] T cells are also called T lymphocytes, one of human white blood cells, originated from bone marrow hematopoietic stem cells, matures in the thymus, then moved to human blood, lymphoid tissue organs, and play immune function. Its role is equivalent to the "warrior" in the human body, which can resist and destroy "enemy" such as infection, tumor, foreign foreign matter. [0003] As the T lymphocyte tumor immune response mechanism is increasingly attached, chimeric antigen receptor T (CAR-T) cell therapy is a new immunotherapy strategy in the field of tumor immunotherapy. In the laboratory, technicians are activated by genetic engineering techniques, and they are attached to the positioning device Car (tumor chimeric antigelastic receptor), and CAR-T cell therapy is a new precisi...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N5/0783
CPCC12N5/0636C12N2500/38C12N2500/32C12N2501/2302C12N2501/2305C12N2501/999C12N2501/515C12N2501/51
Inventor 曾献芳郭威胡玉林
Owner 曾献芳
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