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A-NK cell separating culture method

A technology of separation culture and separation method, which is applied in the direction of tissue culture, animal cells, vertebrate cells, etc., and can solve the problems of time-consuming, labor-intensive, easy-pollution, etc.

Inactive Publication Date: 2005-12-14
TUMOR PREVENTING & TREATING INST HARBIN MEDICALUNIV
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AI Technical Summary

Problems solved by technology

This method is expensive, labor-intensive, time-consuming, easy to pollute, and is not conducive to the development of clinical treatment

Method used

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  • A-NK cell separating culture method
  • A-NK cell separating culture method
  • A-NK cell separating culture method

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] 1. Main reagents and culture medium

[0023] 1. Recombined human interleukin-2 (rhIL-2): product of Nanjing Military Medical Research Institute (100,000 IU / 2ml)

[0024] 2. Recombinant human interleukin 12 (IL-12)

[0025] 3. Lymphocyte separation medium: product of Sigma company

[0026] 4. Phenylalanine methyl ester (PME): a product of Sigma

[0027] 5 MTT (methyl thiazolyldipheny telrazolium bromide): a product of Sigma

[0028] 6. Complete culture medium (CM): adopt RPMI-1640 complete culture medium to cultivate, and it is composed of 90% RPMI-1640 culture medium (Gibco product), 10% human AB serum (provided by the blood station of Harbin Red Cross Center, for human A-NK cell culture) or 10% inactivated fetal bovine serum (Institute of Hematology, Chinese Academy of Medical Sciences, for tumor cell culture), 2mM glutamine, 10mM HEPES, 100u / ml penicillin, 100ug / ml streptomycin.

[0029] 7. Serum-free medium (AIMV): product of Gibco

[0030] Culture conditions ar...

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Abstract

The improved A-NK cell separating culture method is one improved tumor treating biologically therapeutic method, and includes adopting PME to eliminate mononuclear phagocyte without altering phynotype and toxicity of lymphocyte; displacing no-serum culture medium for serum culture medium to obtain similar number and function of the cultured matter and raised clinical safety; combining IL-4 with IL-2 to generate relatively strong stimulation to A-NK cell growth and mature and to raise the killing activity of the activated immunological cell.

Description

technical field [0001] The invention relates to a medical tumor biotherapeutic method technical background [0002] A-NK cells (adherent natural killer cell), also known as A-LAK (adherent Lymphokin Activitate killer cell) cells, have become the focus of adoptive immunotherapy for tumors in recent years due to their strong in vitro proliferation and anti-tumor ability and no killing effect on normal cells. one of the research hotspots. In clinical applications, the complete culture medium commonly used for cell culture currently contains human AB serum. Although it has been tested for various viruses, it may still spread other diseases. The safety is low, and there are batch-to-batch differences, so it is not suitable for quality control. , the price is also very expensive. The serum-free medium that has been released in recent years can meet the requirements of biological cleanliness, has clear components, stable quality, and is convenient for quality control. It overcome...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
Inventor 王志华
Owner TUMOR PREVENTING & TREATING INST HARBIN MEDICALUNIV
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