Separation method and separation kit for neutrophil in blood

Abstract

The invention relates to a separation method and a separation kit for neutrophil in blood, and belongs to the technical field of neutrophil separation. The invention provides the separation method for the neutrophil in blood, which comprises the following steps: sequentially adding a neutrophile granulocyte separating medium, a lymphocyte separating medium and whole blood into a centrifugal tube, enabling the lymphocyte separating medium to be positioned above the neutrophile granulocyte separating medium and the whole blood to be positioned above the lymphocyte separating medium, then centrifuging, and separating to obtain the neutrophile granulocytes. The invention further provides a separation kit for the neutrophil in the blood. The separation kit contains a neutrophil separation medium and a lymphocyte separation medium which are independently packaged respectively. By adopting the separation method or the separation kit provided by the invention, the neutrophil with the purity of about 80% can be rapidly obtained through one-time centrifugation, and the neutrophil and the red blood cells can be separated at the same time through one-time centrifugation, so that the separation method or the separation kit has a very good practical value.

Description

technical field [0001] The invention relates to a separation method and a separation kit for neutrophils in blood, belonging to the technical field of neutrophil separation. Background technique [0002] Neutrophils play a very important role in the body's non-specific immune defense response, and are important phagocytes for the body to resist microbial pathogens, especially the invasion of suppurative bacteria. Peripheral blood leukocytes are mainly neutrophils, which account for 50% to 70% of human peripheral blood leukocytes under normal circumstances. In acute infection or inflammation, extensive tissue damage or necrosis, acute hemolysis and blood loss, acute poisoning, and malignant tumors, the number of peripheral blood neutrophils increases. At present, there have been reported methods for separating human and animal neutrophils, most of which use a single separation medium such as Percoll, Ficoll, etc. to obtain the diluted peripheral blood by gradient centrifugat...

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Problems solved by technology

However, the above-mentioned gradient centrifugation method has very strict requirements on the concentration of the separation liquid and the meticulousness of the operation. Small changes can easily lead to cell separation failure, and it takes a long time. The purity of the cells obtained after centrifugation varies greatly, and there are usually more impurity cells.

Some companies on the market provide commercial neutrophil separation reagents, but the separation effect is still affected by different experimental factors, especially after centrifugation, the cell stratification is not clear, and there are mixed cells, which affects the purity of the target cells. (like figure 1 shown)

In addition, it has also been reported that the method of obtaining neutrophils with higher purity by repeated centrifugation, but increasing the number of centrifugation and too long centrifugation time will cause certain damage to the cells

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