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Method for extracting immune cells from peripheral blood

An immune cell and peripheral blood technology, applied in the direction of blood/immune system cells, animal cells, vertebrate cells, etc., can solve the problems of survival rate, cytotoxicity, single cell damage, etc., and achieve the effect of reducing immune rejection.

Inactive Publication Date: 2018-01-30
重庆斯德姆生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, cryopreservation technology is mainly used to preserve single cells in peripheral blood, which can be revived and induced to culture into various immune cells when needed. The thermodynamic, chemical and physical environment of cells is significantly changed during cryopreservation, accompanied by the risk of biological damage
[0003] In addition, the separation liquid used in the prior art has certain toxicity to the cells, which is not conducive to clinical application, and the survival rate of cells after cryopreservation and recovery may be affected by the processes of cell collection, separation, and cryopreservation

Method used

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  • Method for extracting immune cells from peripheral blood
  • Method for extracting immune cells from peripheral blood

Examples

Experimental program
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Effect test

Embodiment 1

[0024] A method for extracting immune cells from peripheral blood, comprising the steps of:

[0025] (1) Collect human peripheral blood, add anticoagulant, then move it into a centrifuge tube, centrifuge, absorb the upper layer of plasma for later use, add physiological saline to the lower layer of cells, and mix to obtain a cell suspension; wherein, the volume ratio of peripheral blood to normal saline is 1:2;

[0026] (2) Place the cell suspension in the separation medium, centrifuge at 800g for 30min, absorb buffy coat cells to obtain immune cells;

[0027] Wherein, the separation liquid includes the following components by weight: 3 parts of trehalose, 1 part of dextran (Dextran70), 1 part of stephine, 1 part of tremella polysaccharide, 0.5 part of laminarin, and 0.2 part of ultra-low viscosity sodium alginate and 0.1 part of polydiallyldimethylammonium chloride;

[0028] The separation liquid is prepared by the following method: mixing the components by weight, then dis...

Embodiment 2

[0033] A method for extracting immune cells from peripheral blood, comprising the steps of:

[0034] (1) Collect human peripheral blood, add anticoagulant, then move it into a centrifuge tube, centrifuge, absorb the upper layer of plasma for later use, add physiological saline to the lower layer of cells, and mix to obtain a cell suspension; wherein, the volume ratio of peripheral blood to normal saline is 1:4;

[0035] (2) Place the cell suspension in the separation medium, centrifuge at 800g for 30min, absorb buffy coat cells to obtain immune cells;

[0036] Wherein, the separation liquid includes the following components by weight: 6 parts of trehalose, 3 parts of dextran (Dextran70), 3 parts of stephine, 2 parts of tremella polysaccharide, 1.2 parts of laminarin, and 0.5 parts of ultra-low viscosity sodium alginate And 0.2 part of polydiallyl dimethyl ammonium chloride;

[0037] The separation liquid is prepared by the following method: mixing the components by weight, t...

Embodiment 3

[0042] A method for extracting immune cells from peripheral blood, comprising the steps of:

[0043] (1) Collect human peripheral blood, add anticoagulant, then move it into a centrifuge tube, centrifuge, absorb the upper layer of plasma for later use, add physiological saline to the lower layer of cells, and mix to obtain a cell suspension; wherein, the volume ratio of peripheral blood to normal saline is 1:3;

[0044] (2) Place the cell suspension in the separation medium, centrifuge at 800g for 30min, absorb buffy coat cells to obtain immune cells;

[0045] Wherein, the separation liquid includes the following components by weight: 4 parts of trehalose, 1.5 parts of dextran (Dextran70), 1.5 parts of stephine, 1.5 parts of tremella polysaccharide, 0.7 part of laminarin, and 0.3 part of ultra-low viscosity sodium alginate And 0.13 parts of polydiallyl dimethyl ammonium chloride;

[0046] The separation liquid is prepared by the following method: mixing the components by weigh...

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Abstract

The invention discloses a method for extracting immune cells from peripheral blood. The method comprises the following steps: (1) collecting human peripheral blood and adding an anticoagulant; then transferring the mixture into a centrifugal tube and centrifuging; sucking upper-layer blood plasma for later use and adding normal saline into lower-layer cells; mixing to obtain a cell suspension solution; (2) putting the cell suspension solution into a separation solution and carrying out 600g to 800g centrifuging for 20min to 30min; sucking albuginea cells to obtain immune cells; (3) adding theimmune cells with the same volume into cryopreservation liquid and sub-packaging the mixture into cell cryopreservation tubes; then putting the cryopreservation tubes into a program cryopreservation box; cooling to -20 DEG C to -25 DEG C at the speed of 1 DEG C / min to 2 DEG C / min; then cooling to -40 DEG C to -45 DEG C at the speed of 3 DEG C / min to 5 DEG C / min; then cooling to -78 DEG C to -82 DEG C at the speed of 5 DEG C / min to 6 DEG C / min; finally, transferring the cryopreservation tubes into liquid nitrogen and cryopreserving. By adopting the method disclosed by the invention, the separation rate of the immune cells and the survival rate of recovered cryopreserved cells can be effectively improved; the safety of the cells, which are infused back to patients, is improved.

Description

technical field [0001] The invention belongs to the technical field of extracting immune cells, in particular to a method for extracting immune cells from peripheral blood. Background technique [0002] Extracting immune cells from human peripheral blood not only has good prospects in the treatment of cancer, but also from the perspective of health preservation, immune cells can also be extracted from the peripheral blood of patients when they are healthy, and can be used in patients in the future. When I have low immunity or other symptoms, I can infuse the stored immune cells back to the patient to improve the patient's immunity. At present, cryopreservation technology is mainly used to preserve single cells in peripheral blood, which can be revived and induced to culture into various immune cells when needed. The thermodynamic, chemical and physical environment of cells will be significantly changed during cryopreservation, accompanied by the risk of biological damage. ...

Claims

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Application Information

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IPC IPC(8): C12N5/078A01N1/02
Inventor 胡小东王健赵翔熊华强宋彩霞王仁杰邱阳黄启程李国焱唐玲叶劲涛曹玉昊
Owner 重庆斯德姆生物技术有限公司
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