Method for separating and purifying T lymphocytes

A technology for separation and purification of lymphocytes, applied in the field of separation and purification of T lymphocytes, can solve problems such as limited use, unfavorable effective use of T lymphocytes, limited source of umbilical cord blood, etc., and achieve the effect of optimizing extraction conditions

Inactive Publication Date: 2019-04-26
SHENZHEN PREGENE BIOPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to limited sources of umbilical cord blood and limited use due to individual differences, it is not conducive to the effective use of T lymphocytes

Method used

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  • Method for separating and purifying T lymphocytes
  • Method for separating and purifying T lymphocytes
  • Method for separating and purifying T lymphocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A method for separating and purifying T lymphocytes, comprising the steps of:

[0021] 1) Plasma separation: Take whole blood and put it into a centrifuge tube, add anticoagulant, centrifuge at 800g, 4°C for 15 minutes, draw plasma into another centrifuge tube, extinguish the fire at 56°C for 30 minutes, centrifuge for 30 minutes under the same conditions, remove Precipitate, collect supernatant;

[0022] 2) Separation of lymphocytes: dilute whole blood with 0.9% NaCl injection in a clean test tube, mix well; draw 5ml of supernatant into another clean centrifuge tube, add separation solution, and level the diluted whole blood Spread it above the liquid surface of the separation liquid, centrifuge at 300g, 20°C for 30min, and a buffy coat layer appears;

[0023] 3) Washing of lymphocytes: absorb the buffy coat into a centrifuge tube, add 0.9% NaCl injection to dilute, mix well, centrifuge at 400g, 20°C for 10min, discard the supernatant, and repeat the operation once; ...

Embodiment 2

[0026] A method for separating and purifying T lymphocytes, comprising the steps of:

[0027] 1) Plasma separation: Take whole blood and put it into a centrifuge tube, add anticoagulant, centrifuge at 800g, 4°C for 15 minutes, draw plasma into another centrifuge tube, extinguish the fire at 56°C for 30 minutes, centrifuge for 30 minutes under the same conditions, remove Precipitate, collect supernatant;

[0028] 2) Separation of lymphocytes: dilute whole blood with 0.9% NaCl injection in a clean test tube, mix well; draw 5ml of supernatant into another clean centrifuge tube, add separation solution, and level the diluted whole blood Spread it above the liquid surface of the separation liquid, centrifuge at 400g, 20°C for 30min, and a buffy coat layer appears;

[0029] 3) Washing of lymphocytes: absorb the buffy coat into a centrifuge tube, add 0.9% NaCl injection to dilute, mix well, centrifuge at 450g, 20°C for 10min, discard the supernatant, and repeat the operation once; ...

Embodiment 3

[0032] A method for separating and purifying T lymphocytes, comprising the steps of:

[0033] 1) Plasma separation: Take whole blood and put it into a centrifuge tube, add anticoagulant, centrifuge at 800g, 4°C for 15 minutes, draw plasma into another centrifuge tube, extinguish the fire at 56°C for 30 minutes, centrifuge for 30 minutes under the same conditions, remove Precipitate, collect supernatant;

[0034] 2) Separation of lymphocytes: dilute whole blood with 0.9% NaCl injection in a clean test tube, mix well; draw 5ml of supernatant into another clean centrifuge tube, add separation solution, and level the diluted whole blood Spread it above the liquid surface of the separation liquid, centrifuge at 500g, 20°C for 30min, and a buffy coat layer appears;

[0035] 3) Washing of lymphocytes: absorb the buffy coat into a centrifuge tube, add 0.9% NaCl injection to dilute, mix well, centrifuge at 500g, 20°C for 10min, discard the supernatant, and repeat the operation once; ...

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Abstract

The invention discloses a method for separating and purifying T lymphocytes. The method comprises the following four steps: 1) separating plasma; 2) separating lymphocytes; 3) cleaning lymphocytes; 4)purifying lymphocytes. According to the technical scheme of the invention, extraction conditions of T lymphocytes are optimized and effective utilization of T lymphocytes is benefited.

Description

technical field [0001] The invention relates to the technical field of cell extraction and separation, in particular to a method for separating and purifying T lymphocytes. Background technique [0002] T cells are one of the two major lymphocytes in the human body and play an extremely important role in specific cellular immunity. Mature T cells settle in the thymus-dependent area of ​​the peripheral immune organs of the human body, and can circulate through lymphatic vessels, peripheral blood, and interstitial fluid to exert cellular immunity and immune regulation functions. Because T cells have extremely strong specificity and good killing effect in cellular immunity, T cells have broad application prospects in basic immunology research and in the biopharmaceutical industry related to immunotherapy. Usually, in order to obtain a better extraction effect, T cells are separated and extracted by collecting umbilical cord blood. Due to the limited source of umbilical cord b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2509/00
Inventor 张继帅宋宗培蔡清华郑孟韬苏红昌李华秀包朝乐萌
Owner SHENZHEN PREGENE BIOPHARMA CO LTD
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