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Monocyte separating method

A mononuclear cell and cell technology, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of cumbersome operation and relatively high operator proficiency requirements, and achieve the effect of prolonging storage time and preventing impact

Inactive Publication Date: 2010-03-03
CHENGDU RICH SCI IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] This method is cumbersome to operate and requires relatively high operator proficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Through vacuum blood collection technology, a certain amount of whole blood is collected into a test tube, and the blood is mixed with the cell maintenance solution. The container is then put into a centrifuge, and centrifuged for 8 minutes with a relative centrifugal force RCF 800g. Due to the density of PBMC (the density described herein is relative density, the density of pure water at a certain temperature is regarded as 1, and the density of another substance in The density obtained by dividing it under the same conditions, the same below) is different from the density of other components and reagents in the blood. The density of red blood cells is 1.092, the density of separation gel is 1.08, the density of separation liquid is 1.077, and the density of PBMC is 1.07. Therefore, according to the principle of density gradient, zones of different levels are formed: the lowermost layer is the red blood cell layer, followed by the separation gel, separation liquid, mono...

Embodiment 2

[0034] Through vacuum blood collection technology, a certain amount of whole blood is collected into a test tube, and the blood is mixed with the cell maintenance solution. Then with the relative centrifugal force RCF 1200g, the container is put into the centrifuge and centrifuged for 12 minutes, due to the density of PBMC (the densities described in this article are all relative densities, the density of pure water at a certain temperature is regarded as 1, and another substance in The density obtained by dividing it under the same conditions, the same below) is different from the density of other components and reagents in the blood. The density of red blood cells is 1.092, the density of separation gel is 1.08, the density of separation liquid is 1.077, and the density of PBMC is 1.07. Therefore, according to the principle of density gradient, zones of different levels are formed: the lowermost layer is the red blood cell layer, followed by the separation gel, separation liq...

Embodiment 3

[0036] Through vacuum blood collection technology, a certain amount of whole blood is collected into a test tube, and the blood is mixed with the cell maintenance solution. Then with the relative centrifugal force RCF 1000g, the container is put into the centrifuge and centrifuged for 10 minutes, due to the density of PBMC (the densities described in this paper are all relative densities, the density of pure water at a certain temperature is regarded as 1, and another substance in The density obtained by dividing it under the same conditions, the same below) is different from the density of other components and reagents in the blood. The density of red blood cells is 1.092, the density of separation gel is 1.08, the density of separation liquid is 1.077, and the density of PBMC is 1.07. Therefore, according to the principle of density gradient, zones of different levels are formed: the lowermost layer is the red blood cell layer, followed by the separation gel, separation liqui...

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PUM

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Abstract

The invention relates to a separating method for monocytes. The separating method comprises the following steps sequentially: a. injecting a monocyte separating reagent into a vessel, wherein the monocyte separating reagent comprises separating liquid, separating glue and cell maintenance liquid; b. collecting blood into the vessel and causing the blood to be uniformly mixed with the cell maintenance liquid; c. exposing the vessel to the influence of a centrifugal force so that differently-hierarchical zones are formed in the solution in the vessel; and d. sucking out a monocyte layer. The separating method has the advantages that the monocyte separation method is simplified, the blood is directly collected into a test tube, in which the separating reagent is positioned in advance, no anti-freezing blood needs to be prepared or diluted, the cell maintenance liquid prolongs the sample storage time before analysis, the separating glue effectively isolates erythrocytes after the centrifugation, and the mass control is easy.

Description

Technical field: [0001] The present invention relates to a method for isolating monocytes. Background technique: [0002] Immune cells refer to cells that participate in or are related to the immune response, mainly including lymphocytes, monocyte-macrophages, dendritic cells, various granulocytes, and mast cells. According to the unique surface markers and special functions of various types of immune cells, it is an important means to observe the immune state of the body to measure the number and function of various types of immune cells by in vitro or in vivo methods. Peripheral blood mononuclear cells (PBMC) mainly refer to lymphocytes, which are the most commonly used cell population in immunological experiments. At present, in vitro methods are mainly used. The premise of using in vitro methods to detect the number and function of certain cells is to effectively separate the cells from blood components. The traditional separation method is to use a separation liquid m...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/08
Inventor 王军
Owner CHENGDU RICH SCI IND
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