High-efficiency amplification culture solution for tumor infiltration lymphocytes

A lymphocyte, expansion and culture technology, applied in the field of high-efficiency expansion culture medium, can solve the problems of limited application, low expansion efficiency of TIL cells, inability to effectively inhibit the suppressive function of T lymphocytes, etc. Extremely strong tumoricidal effect, inhibitory effect

Inactive Publication Date: 2013-10-30
SHANGHAI NUOHAOYA MEDICAL DIAGNOSTICS TECH CO LTD
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Problems solved by technology

[0006] However, the above studies all have the problems of low expansion efficiency of TIL cells and inability to effectively inhibit the inhibitory function of Treg lymphocytes on cytotoxic T lymphocytes, which limits its application in tumor therapy.

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  • High-efficiency amplification culture solution for tumor infiltration lymphocytes

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Embodiment Construction

[0014] The specific technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and examples, so that those skilled in the art can understand the present invention without constituting a limitation on their rights.

[0015] 1. Clean the surgically resected tumor with HBSS first, then immediately remove necrotic tissue, adipose tissue and obvious normal tissue on the tumor surface under aseptic conditions, rinse with HBSS and cut the tumor into about 0.5mmx0.5mmx0.5mm Tissue fragments, suspended in enzyme digestion solution (RPMI1640 medium containing 0.01% type V hyaluronidase, 0.1% type IV collagenase, and 0.002% type I deoxyribonuclease without AB serum) Digest with shaking and stirring in a water bath at 37°C for 4-5 hours, pass through a mesh, and make a single-cell suspension.

[0016] 2. Place equal volumes of 100% and 75% lymphocyte separation fluids in centrifuge tubes successively, then slowly add the cell...

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Abstract

The invention provides a high-efficiency amplification culture solution for tumor infiltration lymphocytes. The culture solution contains IL-2, IL-12, anti-CD28 and anti-CTLA-4. Compared with the prior art, the high-efficiency amplification culture solution has the advantages that a combined serum-free culture medium containing IL-2, IL-12, anti-CD28, anti-CTLA-4 and other cell factors is adopted, the amplification efficiency of TIL cells is obviously improved, and the amplification efficiency is about 3-10 times higher than the amplification efficiency of a common amplification culture solution; moreover, an inhibitory function of Treg lymphocytes on cytotoxic T lymphocytes can be effectively achieved, and the amplified lymphocyte has an extremely strong tumor killing effect.

Description

technical field [0001] The invention relates to a culture medium for tumor biological immunotherapy, in particular to a high-efficiency amplification culture medium for biological immunotherapy using tumor infiltrating lymphocytes. Background technique [0002] Tumor-infiltrating lymphocytes (TIL) are a group of CTL cells infiltrating tumor sites that have been activated by tumor antigens. It was pioneered by Rosenberg et al. in 1986. The specific method is to isolate TIL cells from tumor tissue, pleural effusion or peripheral blood of patients, amplify them in vitro with culture medium containing IL-2 and CD3 antibodies to a certain amount, and then infuse them back to the patient (Belldegrun A , MuulL and Rosenberg S. Interleukin 2 Expanded Tumor-infiltrating Lymphocytes in Human Renal Cell Cancer: Isolation, Characterization, and Antitumor Activity. Cancer Res 1988; 48: 206-214.). The expanded TIL cells are mainly mature T cells, CD3+ cells can reach more than 95%, and t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 方存磊
Owner SHANGHAI NUOHAOYA MEDICAL DIAGNOSTICS TECH CO LTD
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