Separation method for hepatic inherent and infiltrating macrophages

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...

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 based on the expression of monocyte-macrophage surface markers F4 / 80 and CD11b, but macrophages are a complex group that may exist in both infiltrating and intrinsic macrophages Cells with high or low expression of F4 / 80 and CD11b, so these two markers cannot accurately distinguish macrophage origin

Images