PBMC (peripheral blood mononuclear cell) lymphocyte rapid extraction method

A technology of buffy coat and extraction method, which is applied in the field of PBMC buffy coat rapid extraction method, can solve problems such as difficulty in ensuring the quality of buffy coat, difficulty in extracting buffy coat, and increased experimental costs, so as to shorten the centrifugation time and reduce the cost , the effect of simple method

Inactive Publication Date: 2017-02-15
SINOBIOWAY CELL THERAPY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Looking at the above methods, it can be found that the current PBMC buffy coat extraction methods have many shortcomings such as long experiment time and difficulty in extracting buffy coat, and the quality of buffy coat is difficult to guarantee. However, if a special sieve plate or a special centrifuge tube is used to simplify the The experimental operation shortens the experimental time and increases the additional experimental cost

Method used

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  • PBMC (peripheral blood mononuclear cell) lymphocyte rapid extraction method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The PBMC buffy coat quick extraction method comprises the following steps:

[0025] S1, extract buffy coat from whole blood;

[0026] S2. Add the sallow layer to the lymph separation liquid for centrifugation, the centrifugal speed is 1500rpm, the centrifugal force speed-up is set to 80% braking, and the centrifugal force deceleration is set to 50% braking;

[0027] S3. After centrifuging for 20 minutes, the buffy coat was extracted.

Embodiment 2

[0029] The PBMC buffy coat quick extraction method comprises the following steps:

[0030] S1, extract buffy coat from whole blood;

[0031] S2. Add the sallow layer to the lymph separation liquid for centrifugation, the centrifugal speed is 2000rpm, the centrifugal force speed-up is set to 50% braking, and the centrifugal force deceleration is set to 80% braking;

[0032] S3, after centrifuging for 15 minutes, the buffy coat was extracted.

Embodiment 3

[0034] The PBMC buffy coat quick extraction method comprises the following steps:

[0035] S1, centrifuging the whole blood to extract the buffy coat;

[0036] S2, adding the sallow layer to the lymph separation liquid for centrifugation, the centrifugation speed is 1600rpm, the speed up and down are both set to 70% braking, and the centrifuge used for centrifugation is a Thermo ST40R centrifuge;

[0037] S3. After centrifuging for 18 minutes, the buffy coat was extracted.

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Abstract

The invention discloses a PBMC (peripheral blood mononuclear cell) lymphocyte rapid extraction method, which comprises the following steps: extracting a buffy coat from whole blood; adding the buffy coat to a lymphocyte separating solution and conducting centrifuging at a centrifugal rotational speed of 1500-2000rpm, setting centrifugal force speed raising as being 50-80% braked and setting centrifugal force speed reducing as being 50-80% braked; and conducting centrifuging for 15-20min and extracting a buffy coat. The prepared mononuclear lymphocyte product provided by the invention is good in quality and high in extraction efficiency, and the method is simple, short in consumed time, low in amount of consumables and low in cost.

Description

technical field [0001] The invention relates to the technical field of PBMC buffy coat extraction, in particular to a rapid extraction method of PBMC buffy coat. Background technique [0002] The current PBMC experimental method mainly adopts manual operation with timing and constant speed centrifugation, or assists some newly developed special screens and centrifuge tubes for operation. The extraction of mononuclear lymphocytes (commonly known as buffy coat) in the PBMC test method is particularly critical. However, the commonly used experimental methods generally have a long centrifugation time, and the auxiliary special consumables increase additional experimental costs, making the experimental operation of the PBMC test method difficult. Difficult. [0003] Among the current existing methods, the application number CN201510768205.9 "A Method for Separating PBMCs" discloses that the buffy coat is extracted by centrifugation for 20-30 minutes under the conditions of 15-18...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2509/00
Inventor 钱鹏赵依妮许国贞刘振云
Owner SINOBIOWAY CELL THERAPY CO LTD
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