Method for extracting and collecting peripheral blood mononuclear cells

A technology for nuclear cells and peripheral blood, applied in the field of extraction and collection of peripheral blood mononuclear cells, can solve the problems of unfavorable application, unstable cell yield and activity rate, etc., and achieve the effect of high activity and high cell yield

Pending Publication Date: 2021-03-09
海南优尼科尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are many methods for the extraction and separation of related PBMCs, but the cell yield and activity rate are unstable, which is not conducive to its application to a certain extent.

Method used

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  • Method for extracting and collecting peripheral blood mononuclear cells

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Effect test

Embodiment 1

[0021] A method for extracting and collecting peripheral blood mononuclear cells, comprising the following steps:

[0022] 1) Peripheral blood collection: Collect 100ml of human peripheral blood in a blood collection bag containing sodium citrate anticoagulant. After the collection is completed, the blood collection bag is refrigerated and transported in a sealed sterile transport box at a temperature of 4-8°C;

[0023] 2) Peripheral blood pretreatment: sterilize the blood collection bag containing peripheral blood transported in refrigerated storage in step 1), then transfer the peripheral blood in the blood collection bag to a 50ml centrifuge tube, take samples for counting, and perform corresponding detection;

[0024] 3) Centrifugation: Place the centrifuge tube containing the peripheral blood described in step 2) in a centrifuge, centrifuge at 2500 rpm for 10 minutes at 4°C, adjust the centrifuge to increase speed by 2 and decrease speed by 1, and the blood in the centrifu...

Embodiment 2

[0032] A method for extracting and collecting peripheral blood mononuclear cells, comprising the following steps:

[0033] 1) Peripheral blood collection: Collect 50ml of human peripheral blood in a blood collection bag containing sodium citrate anticoagulant. After the collection is completed, the blood collection bag is refrigerated in a sealed sterile transport box at a temperature of 4-8°C;

[0034] 2) Peripheral blood pretreatment: sterilize the blood collection bag containing peripheral blood transported in refrigerated storage in step 1), then transfer the peripheral blood in the blood collection bag to a 50ml centrifuge tube, take samples for counting, and perform corresponding detection;

[0035] 3) Centrifugation: place the centrifuge tube containing the peripheral blood described in step 2) in a centrifuge, and centrifuge at 2100rpm for 15min at 4°C. Respectively, the upper layer of plasma and the lower layer of blood cells;

[0036] 4) Plasma extraction: Aspirate ...

Embodiment 3

[0043] A method for extracting and collecting peripheral blood mononuclear cells, comprising the following steps:

[0044] 1) Peripheral blood collection: Collect 80ml of human peripheral blood in a blood collection bag containing sodium citrate anticoagulant. After the collection is completed, the blood collection bag is refrigerated and transported in a sealed sterile transport box, and the refrigerated temperature is 4-8 °C;

[0045] 2) Peripheral blood pretreatment: Sterilize the blood collection bag containing peripheral blood transported by refrigeration in step 1), then transfer the peripheral blood in the blood collection bag to a 50ml centrifuge tube, take samples for counting, and do corresponding detection;

[0046] 3) Centrifugation: the centrifuge tube containing peripheral blood described in step 2) was placed in a centrifuge, centrifuged at 2500rpm for 10min at 4°C, the centrifuge was adjusted to increase speed 2, decrease speed 1, and the blood in the centrifuge...

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Abstract

The invention discloses a method for extracting and collecting peripheral blood mononuclear cells. The method comprises the following steps: 1) collecting peripheral blood; 2) pretreating peripheral blood; 3) performing centrifuging; 4) extracting plasma; 5) performing plasma inactivation; 6) collecting mononuclear cell layers; 7) centrifuging the mononuclear cell layers; 8) performing cleaning; and 9) extracting mononuclear cells. The invention belongs to the technical field of cell extraction, and particularly provides a method for separating peripheral blood mononuclear cells, which is usedfor obtaining the peripheral blood mononuclear cells by centrifugal extraction of Ficoll lymphocyte separation liquid and is high in cell yield and high in activity.

Description

technical field [0001] The invention belongs to the technical field of cell extraction, and specifically refers to a method for extracting and collecting peripheral blood mononuclear cells. Background technique [0002] Peripheral blood mononuclear cells are cells with a single nucleus in peripheral blood, mainly including monocytes, lymphocytes (B cells and T cells), phagocytes, dendritic cells and NK cells, etc. These cells are the body's immune response important cells. Peripheral blood mononuclear cells are rich in peripheral blood stem cells, which can treat metabolic genetic diseases, tumor diseases, immunodeficiency diseases, malignant blood diseases, etc. [0003] In recent years, immunotherapy, as the fourth mode of comprehensive tumor treatment, has been more and more widely used. Studies have found that peripheral blood mononuclear cells can proliferate and differentiate in vitro for the treatment of diseases. In addition, peripheral blood mononuclear cells are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/10C12N2509/00
Inventor 王晓宇林冠妃郭康合李崴蔡开婷张万程李秋宛王岱桑
Owner 海南优尼科尔生物科技有限公司
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