Isolated culture method of rat liver sinusoidal endothelial cells

Abstract

The invention discloses an isolated culture method of rat liver sinusoidal endothelial cells. In the separation step, a biopsy needle is pioneered to be used for creating a digestive cavity, and the digestive cavity which is completely in the liver is formed, so that digestion is stable and efficient, and the contamination rate is low. The isolated culture method uses a combination of multiple enzymes instead of using pancreatin, thus protecting cells to the utmost extent; furthermore, components such as endothelial factor and heparin which are not beneficial to the growth of liver parenchyma cells, but are beneficial to the growth of liver sinusoidal endothelial cells are added into digestive juice, and the screening of the parenchyma cells and the liver sinusoidal endothelial cells is started in the cell separation step. The isolated culture method does not need blowing and beating from beginning to end, thus protecting the vitality of the liver sinusoidal endothelial cells to the utmost extent, and the cultured cells are high in cell viability and good in activity. The isolated culture method has greater promotion significance, and provides a good cell culture solution for related research on the basis of liver sinusoidal endothelial cell culture.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for separating and culturing rat liver sinusoidal endothelial cells. Background technique [0002] Endothelial cells participate in a variety of physiological functions and play an important role in many pathological processes. The liver contains two distinct endothelial cell types: vascular endothelium and hepatic sinusoidal endothelium. Hepatic sinusoidal endothelial cells are a type of microvascular endothelial cells with a unique phenotype, which is similar to dendritic cells, and a unique function, which is to present antigens to CD4+ T cells. Hepatic sinusoidal endothelial cells thus represent a novel, "non-professional" antigen-presenting cell within the organ, participate in local immune responses within the organ, and mediate immune tolerance within the liver. The microenvironment system in the liver, including some soluble substances released by hepatic ...

AI Technical Summary

Problems solved by technology

The collagenase perfusion method, no matter the traditional in situ perfusion method, two-step perfusion method, or even the newly improved three-step perfusion method, has a better effect on the separation of liver parenchymal cells, but it still has the problem of excessive digestion of hepatic sinusoidal endothelial cells. , and in the process of separation, most of the liver parenchymal cells are still present, so it is very difficult to screen and isolate liver sinusoidal endothelial cells with high purity

At the same time, it should be noted that during the separation process of the above two methods, the cells need to be repeatedly blown and blown to completely separate the cells, which will cause further damage to the liver sinusoidal endothelial cells, resulting in a further decrease in the yield and activity of the cells. decline

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