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Experiment method for efficiently amplifying NK (Natural Killer) cells of human peripheral blood

A technology of NK cells and human peripheral blood, applied in the biological field, can solve the problems of low cell number expansion and low NK cell phenotype, and achieve the effect of high lymphatic group, high number and high viability

Inactive Publication Date: 2018-03-09
TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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  • Abstract
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  • Application Information

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Problems solved by technology

[0003] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an experimental method for efficiently expanding human peripheral blood NK cells. The present invention uses corresponding technical means to remove miscellaneous cells, and uses factor compounding to remove them within a suitable time. Factors solve the problem of low phenotype of NK cells obtained after traditional factor stimulation and low cell number expansion multiples. Through short-term culture, NK cells with high lymphatic population, high activity rate and high number can be obtained

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  • Experiment method for efficiently amplifying NK (Natural Killer) cells of human peripheral blood
  • Experiment method for efficiently amplifying NK (Natural Killer) cells of human peripheral blood
  • Experiment method for efficiently amplifying NK (Natural Killer) cells of human peripheral blood

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Embodiment Construction

[0024] The present invention will be further described in detail below in conjunction with the accompanying drawings and through specific embodiments. The following embodiments are only descriptive, not restrictive, and cannot limit the protection scope of the present invention.

[0025] An experimental method for efficiently expanding human peripheral blood NK cells, the steps are as follows:

[0026] (1) Centrifuge the peripheral blood at 800g for 10min (8 liters and 4 drops), take the supernatant to inactivate at 56°C for 30 minutes, centrifuge at 2000g for 10 minutes (8 liters and 8 drops), and take the supernatant as autologous serum;

[0027] (2) Dilute the blood twice with normal saline, slowly add the resuspended liquid into the centrifuge tube containing the ficoll separation medium, the ratio of Ficoll to liquid = 3:4 (volume ratio), (15ml ficoll, 20ml liquid) centrifuge at 600g , 40min (1 liter 0 drop);

[0028] (3) Aspirate the buffy coat, rinse with normal saline...

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Abstract

The invention relates to an experiment method for efficiently amplifying NK (Natural Killer) cells of human peripheral blood. Parenchyma cells are removed by adopting a corresponding technical means;factors are used for compounding and the factors are removed within proper time; the problems that phenotypes of the NK cells obtained by stimulating traditional factors are not high and the amplification times of cell quantity are few are solved; the NK cells with high lymphocyte groups, high viability and high quantity can be obtained through short-time culture.

Description

technical field [0001] The invention is used in the field of biotechnology, mainly for cell culture and immunotherapy, and relates to a method for cultivating NK cells with high purity and high expansion rate in vitro by using mononuclear cells in autologous peripheral blood. Background technique [0002] NK cells (natural killer cel1) have been discovered 35 years ago. They belong to the innate immune system and are the first line of defense of the human immune system. NK cells have no MHC restriction in the process of killing target cells, and can effectively kill a variety of tumor cells. At present, the adoptive therapy of NK cells has achieved a certain clinical effect in the treatment of anti-virus and anti-tumor, but the number of NK cells in the peripheral blood of normal human beings is small, accounting for only 10% to 15% of peripheral blood lymphocytes. Exist in peripheral tissues, such as liver, peritoneal cavity, placenta, etc. Establishing an effective NK ce...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0646C12N2501/2315C12N2501/515C12N2501/599
Inventor 周瑶马洁赵刚冯郸许蕊翟雷垒
Owner TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD