Separation method for hepatic inherent and infiltrating macrophages

A technology of macrophages and separation method, which is applied in the field of separation of resident and infiltrating macrophages in the liver, and can solve the problems of inability to accurately distinguish the source of macrophages, lack of surface markers of liver infiltration and resident macrophages, etc.

Pending Publication Date: 2020-01-10
NANTONG UNIVERSITY
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AI Technical Summary

Problems solved by technology

The respective roles of liver-infiltrating and resident macrophages during liver injury are unclear, but surface markers that can effectively distinguish liver-infiltrating and resident macrophages are lacking.
Existing methods mostly distinguish liver macrophages from different sources base...

Method used

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  • Separation method for hepatic inherent and infiltrating macrophages
  • Separation method for hepatic inherent and infiltrating macrophages
  • Separation method for hepatic inherent and infiltrating macrophages

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Embodiment Construction

[0060] In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, the following will describe in detail with reference to the drawings and specific embodiments.

[0061] The present invention provides a method for separating liver intrinsic and infiltrating macrophages, comprising the following steps:

[0062] (1) Establish and identify a mouse allogeneic bone marrow transplantation model (such as figure 1 shown), the recipient mouse monocyte-macrophage surface marker is CD45.2, and the donor mouse monocyte-macrophage surface marker is CD45.1; the specific steps include:

[0063] a. Preparation of CD45.1 C57BL / 6 mouse bone marrow cells:

[0064] Sacrifice the mice and disinfect the skin;

[0065] The long bones of mouse limbs were freed and soaked in RPMI-1640;

[0066] Cut off the cartilage at both ends and flush out the cells in the bone marrow cavity;

[0067] Repeatedly blow and beat to mix the cel...

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Abstract

The invention provides a separation method for hepatic inherent and infiltrating macrophages. The separation method comprises the following steps of (1) establishing and identifying mouse allogeneicbone marrow transplantation model, wherein a surface marker of a mononuclear macrophage of a recipient mouse is CD45.2, and a surface marker of a mononuclear macrophage of a donor mouse is CD45.1; (2)observing and irradiating the influence on hepatic parenchymal cells and inherent macrophages with an IHC method and a flow cytometry; and (3) irradiating a mononuclear cell of a mouse liver in a transplantation group with primary isolation, and sorting the hepatic inherent and infiltrating macrophages with the flow cytometry according to the surface marker of the mononuclear macrophage of the donor mouse, and describing the quantities, distribution and phenotypes of the hepatic inherent and infiltrating macrophages. In the separation method provided by the invention, by removing the CD45.2 recipient mouse bone marrow derived macrophages by irradiation and supplementing and replacing bone marrow cells of the recipient mouse with CD45.1 donor mouse bone marrows, the bone marrow derived infiltrating macrophages and the bone marrow derived inherent macrophages have different molecular phenotypes and achieves the effect of accurately and efficiently separating different hepatic macrophages.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating liver intrinsic and infiltrating macrophages. Background technique [0002] The liver is not only an important place for substance metabolism, but also an important immune organ. Hepatic macrophages are composed of innate and infiltrating macrophages, with complex phenotypes and functions, and play an important role in maintaining the normal physiological functions of the liver, liver damage caused by various poisons, drugs and pathogenic microorganisms, and various liver metabolic diseases. It has always been an important field in the study of liver pathology. [0003] Liver-intrinsic macrophages and infiltrating macrophages derived from peripheral blood monocytes are cell populations with different origins and different biological functions. In recent years, studies have shown that the number of inherent macrophages decreases in the early stage...

Claims

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Application Information

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IPC IPC(8): C12N5/0786C12N5/077
CPCC12N5/0645C12N5/0652C12N2509/00C12N2501/599
Inventor 季菊玲季煜华沈宓李静何理孙玉风陆鹏吕秀芳
Owner NANTONG UNIVERSITY
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