Rat hepatic stellate cell separation method

Abstract

The invention provides a rat hepatic stellate cell separation method, which comprises the following steps of (a) injecting 10-percent chloral hydrate into the abdominal cavity for anaesthetizing a rat, and laparotomizing the abdominal cavity of the rat so as to expose the liver and the portal vein; (b) performing heparin intravenous injection, and preheating perfusion liquid for liver perfusion; (d) taking the liver subjected to sufficient perfusion, cutting the liver into pieces, and performing oscillation digestion in preheated digestion liquid; (e) filtering digested liver tissues through a multilayer sieve screen; performing low-speed centrifugation; sucking supernatant; performing centrifugation again to obtain cell precipitates; and performing once cleaning by cleaning liquid; (f) resuspending the obtained cell precipitates, and performing Nycoden gradient centrifugal separation; (g) cleaning the cells for many times, resuspending the cell precipitates by a complete culture medium, and performing inoculated culture; and (h) performing cell immunofluorescent identification on rat hepatic stellate cells. The rat hepatic stellate cell separation method provided by the invention can be used for obtaining rat hepatic stellate cells which have high yield, high purity, high activity and can be activated.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for isolating rat hepatic stellate cells. Background technique [0002] Hepatic stellate cells (HSCs) are non-parenchymal cells unique to the liver, accounting for about 8%-13% of the total number of liver cells, 33% of the total number of non-parenchymal cells in the liver, and 1.4% of the total volume of the liver. , is a single layer of adherent growth cells, the cells are star-shaped, spindle-shaped, polygonal, spindle-shaped, with pseudopodia, and the cytoplasm contains many lipid droplets, also known as hepatic fat storage cells. Hepatic stellate cells are the main cell group that synthesizes extracellular matrix in the liver. They can not only secrete proteoglycans, glycoproteins and other extracellular matrix components, but also synthesize a certain amount of collagenase to maintain the normal basement membrane structure. Participate in the microcirculat...

AI Technical Summary

Problems solved by technology

The hepatic stellate cells isolated by the commonly used liver in situ perfusion method are often mixed with Kupffer cells, the amount of stellate cells obtained is small, the activity is low, and the obtained hepatic stellate cells are in a static state and cannot be activated.

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