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Medium for gamma delta T cells amplified in vitro and method

A technology of in vitro expansion and culture medium, which is applied in the field of culture medium for in vitro expansion of γδT cells, can solve the problem of no tumor cell immunotherapy products, etc., and achieve the effect of fast cell growth, easy operation and controllable quality.

Active Publication Date: 2017-02-15
ANHUI PROVINCIAL HOSPITAL
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  • Claims
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Problems solved by technology

At present, my country's specific cell therapy is still in the technical research stage. Although non-specific cell therapy technology has been used in clinical practice for nearly 30 years, there has not been a real tumor cell immunotherapy product yet.

Method used

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  • Medium for gamma delta T cells amplified in vitro and method
  • Medium for gamma delta T cells amplified in vitro and method
  • Medium for gamma delta T cells amplified in vitro and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Expansion of human γδT cells

[0056] 1. Isolation of peripheral blood mononuclear cells (PBMC cells)

[0057] 1. Peripheral blood sample: Take 50ml of peripheral blood from a healthy person or a tumor patient, anticoagulate with heparin, and obtain the informed consent of the blood donor.

[0058] 2. Separation steps:

[0059] (1) Centrifuge the peripheral blood sample at 800g at 25°C for 10 minutes, draw the supernatant (plasma) into a 50ml sterile centrifuge tube, precipitate into peripheral blood cells, place on ice, and use to separate PBMC cells.

[0060] (2) Incubate the supernatant obtained in step (1) in a water bath at 56°C for 30 minutes to inactivate it; then centrifuge at 800g for 10 minutes, draw the supernatant into a 50ml sterile centrifuge tube, which is autologous plasma , stored at -20°C for later use.

[0061] (3) Use sterile PBS solution to dilute the peripheral blood cells obtained in step (1) to 60ml to obtain a peripheral blood cell...

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Abstract

The invention discloses a medium for gamma delta T cells amplified in vitro and a method. The medium is a mammalian cell serum-free medium which successively contains L-glutamine, ribonucleotide, deoxyribonucleoside, blood plasma, human interleukin-2, human interleukin-7, phytohemagglutinin and zoledronic acid, and the concentrations of the various components in the medium are successively 1.45-2.55 mM, 5.5-9.5 mg / L, 5.5-9.5 mg / L, 0.3-0.5% (v / v), 1.0* 105-3.0* 105 IU / L, 5.0-15.0 mu g / L, 0.5-1.5 mg / L and 0.5-1.5 mu mol / L. Human gamma delta T cells are cultured by the medium, and are high in growth speed, large in number, high in purity, high in lethality to tumor cells and stable in activity.

Description

technical field [0001] The invention relates to the field of biotechnology, a culture medium and a method for expanding γδ T cells in vitro. Background technique [0002] Tumor is caused by the body's insufficient immune function to overcome various pathogenic factors, and its occurrence and development are closely related to the immune status of the host. In recent decades, surgery, chemotherapy and radiotherapy have become the main means of clinical treatment of tumors, and have achieved certain therapeutic effects, but the recurrence rate and mortality of tumors are still high, mainly due to tumor cells' response to chemical drugs or radiation. related to the formation of tolerance. The new tumor cell immunotherapy technology can eliminate the remaining tumor cells in patients after surgery, radiotherapy and chemotherapy by restoring and enhancing the immune surveillance ability and immune killing function of tumor patients, so as to prevent tumor recurrence, metastasis ...

Claims

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Application Information

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IPC IPC(8): C12N5/0783A61K35/17A61P35/02A61P35/00
CPCA61K35/17C12N5/0636C12N2500/40C12N2500/42C12N2500/84C12N2501/2302C12N2501/2307C12N2501/24
Inventor 程民胡世莲陈稳沈干沈国栋徐婷娟
Owner ANHUI PROVINCIAL HOSPITAL
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