Method for purifying cord blood mononuclear cells

A technology of nuclear cells and blood cells, applied in the field of biomedicine, can solve the problem of low purity of umbilical cord blood mononuclear cells, and achieve the effects of large number, high purity and high viability.

Inactive Publication Date: 2019-05-03
北京博奥晶典启衡生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For this reason, the embodiment of the present invention provides a method for purifying umbilical cord blood mononuclear cells to solve the problem of low purity of umbilical cord blood mononuclear cells in the prior art

Method used

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  • Method for purifying cord blood mononuclear cells
  • Method for purifying cord blood mononuclear cells
  • Method for purifying cord blood mononuclear cells

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Divide 60ml of umbilical cord blood samples into 50ml centrifuge tubes, centrifuge at 500g for 10min to obtain blood cells and plasma, and collect the plasma for later use.

[0057] Preheat hydroxyethyl starch in a water bath at 30°C for 15 minutes, then add the preheated hydroxyethyl starch to blood cells for precipitation, the volume ratio of blood cell sedimentation to hydroxyethyl starch is 3:1, mix well and let stand 3min, then add normal saline, the volume ratio of normal saline to blood cell sediment is 5:3, then mix and let stand for 30min.

[0058] The cell suspension in the upper layer was taken and centrifuged at a centrifugal force of 500 g for 8 min to obtain lymphocyte precipitates.

[0059] Mix the lymphocyte pellet with the spare plasma to obtain a lymphocyte suspension, and then slowly add it to the Ficoll liquid surface to make a clear interface. The volume ratio of the lymphocyte suspension to the Ficoll liquid surface is 1:1, and then use 400g centri...

Embodiment 2

[0063] Divide 60ml of umbilical cord blood samples into 50ml centrifuge tubes, centrifuge at 500g for 10min to obtain blood cells and plasma, and collect the plasma for later use.

[0064] Preheat hydroxyethyl starch in a water bath at 30°C for 15 minutes, then add the preheated hydroxyethyl starch to blood cells for precipitation, the volume ratio of blood cell sedimentation to hydroxyethyl starch is 3:1, mix well and let stand 3min, then add normal saline, the volume ratio of normal saline to blood cell sediment is 5:3, then mix and let stand for 30min.

[0065] The cell suspension in the upper layer was taken and centrifuged at a centrifugal force of 500 g for 8 min to obtain lymphocyte precipitates.

[0066] Mix the lymphocyte pellet with the spare plasma to obtain a lymphocyte suspension, and then slowly add it to the Ficoll liquid surface to make a clear interface. The volume ratio of the lymphocyte suspension to the Ficoll liquid surface is 1:1, and then use 400g centri...

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Abstract

The embodiment of the invention discloses a method for purifying cord blood mononuclear cells. The method comprises: step 1, centrifuging human umbilical cord blood to obtain blood cells and plasma; step 2, subjecting the blood cells to erythrocyte sedimentation with hydroxyethyl starch to obtain a lymphocyte suspension; and step 3, performing gradient centrifugal separation on the lymphocyte suspension by using a Ficoll reagent so as to obtain the cord blood mononuclear cells. The method can effectively reduce the number of erythrocyte and thrombocyte in the final product, and the number of the purified cord blood mononuclear cells is large, and the purity and the motility rate are high.

Description

technical field [0001] The embodiment of the present invention relates to the technical field of biomedicine, in particular to a method for purifying umbilical cord blood mononuclear cells. Background technique [0002] Whole blood is called whole blood after anticoagulant treatment. Whole blood is mainly divided into three layers after centrifugation, which are plasma layer, buffy coat layer and red blood cell layer from top to bottom. The plasma layer mainly contains plasma, water, protein, salts and various ions; the buffy coat mainly includes platelets, lymphocytes, peripheral blood mononuclear cells and granulocytes. layer; red blood cell layer can be simply divided into young red blood cells and normal red blood cells, because the two are in different growth stages of red blood cells, the specific gravity is slightly different. [0003] Cord blood mononuclear cells (hereinafter referred to as CBMC) refer to cells with a single nucleus in cord blood, including hematopo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
Inventor 沈力坚徐云龙王健侯智勇李明辉代悦杨晓鹏
Owner 北京博奥晶典启衡生物科技有限公司
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