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Method for quickly separating large quantity of PBMCs (peripheral blood mononuclear cells) from human peripheral blood

A technology of human peripheral blood and separation method, which is applied in the field of rapid separation of PBMC from a large number of human peripheral blood, can solve the problems of unfavorable separation of PBMC and cell viability, long operation time, complicated separation method, etc., and achieves easy operation and reduced operation steps. , The effect of saving operation time

Inactive Publication Date: 2016-11-09
SINOBIOWAY CELL THERAPY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is still operable for samples with a small amount of blood, but for a large amount of blood (>100ML), the separation method is cumbersome and the operation time is long, which is not conducive to the separation of PBMCs and the viability of cells

Method used

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  • Method for quickly separating large quantity of PBMCs (peripheral blood mononuclear cells) from human peripheral blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A method for rapidly separating a large amount of human peripheral blood PBMCs, comprising the steps of:

[0021] S1, sending human peripheral blood into a centrifuge tube for centrifugation;

[0022] S2. Absorb the sallow layer, and add physiological saline to it for dilution;

[0023] S3, sending the product obtained in S2 into a centrifuge tube containing lymphocyte separation liquid, wherein the product obtained in S2 is located in the upper layer of the lymphocyte separation liquid;

[0024] S4, centrifugal separation, the middle buffy coat layer was sucked, washed, and the supernatant was discarded by centrifugation to obtain PBMCs.

Embodiment 2

[0026] A method for rapidly separating a large amount of human peripheral blood PBMCs, comprising the steps of:

[0027] S1. Send 30ml of human peripheral blood into a centrifuge tube for centrifugation at a centrifugal speed of 1800rpm and a centrifugation time of 10min;

[0028] S2. After removing the upper layer of plasma, absorb the sallow layer, and add physiological saline to it for dilution;

[0029] S3, sending the product obtained in S2 into a centrifuge tube containing lymphocyte separation liquid, wherein the product obtained in S2 is located in the upper layer of the lymphocyte separation liquid;

[0030] S4, centrifugal separation, the middle buffy coat layer was sucked, washed, and the supernatant was discarded by centrifugation to obtain PBMCs.

Embodiment 3

[0032] A method for rapidly separating a large amount of human peripheral blood PBMCs, comprising the steps of:

[0033] S1. Send 180ml of human peripheral blood into a centrifuge tube for centrifugation at a centrifugation speed of 1800rpm and a centrifugation time of 10min;

[0034] S2. After removing the upper layer of plasma, absorb the sallow layer, and add physiological saline to it for dilution;

[0035] S3, sending the product obtained in S2 into a centrifuge tube containing lymphocyte separation liquid, wherein the product obtained in S2 is located in the upper layer of the lymphocyte separation liquid;

[0036] S4, centrifugal separation, the middle buffy coat layer was sucked, washed, and the supernatant was discarded by centrifugation to obtain PBMCs.

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Abstract

The invention discloses a method for quickly separating a large quantity of PBMCs (peripheral blood mononuclear cells) from human peripheral blood. The method comprises the following steps: the human peripheral blood is transferred to a centrifuge tube for centrifugation; a grey yellow layer is absorbed, and normal saline is added to the layer for dilution; a product is sent to a centrifuge tube containing a lymphocyte separation medium, and the product obtained in the S2 is located on an upper layer of the lymphocyte separation medium; centrifugal separation is performed, an intermediate albugineous coat is absorbed, washed and centrifuged, supernatant is removed, and the PBMCs are obtained. For separation of a large quantity of the PBMCs from the human peripheral blood, the operation steps can be reduced, the operation time and reagent supplies are saved, reagent consumption is reduced, and PBMC separation and cell viability improvement are facilitated.

Description

technical field [0001] The invention relates to the technical field of peripheral blood PBMC separation, in particular to a method for rapidly separating a large number of human peripheral blood PBMCs. Background technique [0002] Autoimmune T cell technology refers to the isolation of T cells from the patient's peripheral blood, through in vitro culture, expansion, preparation, and reinfusion into the patient's body to induce the body's own immune response. At present, T cells are mainly obtained from human peripheral blood PBMC. The main separation method is to directly mix human peripheral blood with PBS in equal amounts, then add lymph separation fluid for gradient separation, and finally obtain PBMC. This method is still operable for samples with a small amount of blood, but for a large amount of blood (>100ML), the separation method is cumbersome and the operation time is long, which is not conducive to the separation of PBMCs and the viability of cells. Contents...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/00
Inventor 钱鹏赵依妮许国贞刘振云
Owner SINOBIOWAY CELL THERAPY CO LTD
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