A method for isolating and culturing hepatic stellate cells of long-clawed gerbils

Abstract

The invention discloses a separating method and culturing method for hepatic stellate cells of a Mongolian gerbil. The separating method includes the steps that 1, the livers of the Mongolian gerbil are moved into a combined enzyme solution to be subjected to perfused digestion, then liver capsules and Glisson pedicles are removed, the livers are smashed to be pasty, the paste is moved into a combined enzyme solution, the mixture is stirred for digestion, and a stirred digestion product is obtained; 2, a first Hank's solution is added, the mixture is filtered, liver tissues are obtained, then a second Hank's solution is added, primary centrifuging is carried out, sediment is obtained, the sediment is suspended again through a third Hank's solution, then a Nycodenz solution is added, the mixture is mixed to be even, centrifuging is carried out again, and a turbid band, namely, the separated hepatic stellate cells is obtained. The separating and culturing method includes the steps of separating and culturing. By means of the methods, the yield and the survival rate of the hepatic stellate cells can be increased, the purity of the HSCs can be guaranteed, and a foundation is laid for further researching of deposition of cell matrixes and forming of fibrosis of the hepatic stellate cells after the livers are injured.

Description

technical field [0001] The invention relates to the field of separating and culturing hepatic stellate cells, in particular to a method for isolating and culturing hepatic stellate cells of gerbils. Background technique [0002] Hepatic stellate cells (HSCs), also known as fat-store cells (FSCs) and interstitial cells, are mainly distributed in the perisinusoidal space between liver cells and hepatic sinusoidal endothelial cells (Disse space). ). HSC plays a central role in the occurrence and development of liver fibrosis, and is the main factor that promotes the development of liver fibrosis. Resting HSCs are mainly involved in the regulation of vitamin A metabolism. When the liver is damaged by chemical, physical and biological factors, HSCs will proliferate and be activated to synthesize collagen-based extracellular matrix (ECM). ) and various cytokines such as MMPs, TNF-α, etc., participate in the repair of damaged liver tissue. When stimulation continues to occur, a ...

AI Technical Summary

Problems solved by technology

This technical scheme adopts the in situ perfusion method to separate HSCs. Because the liver is perfused in vivo, the perfusate is not easy to recover, and the amount of enzyme used is large, and it is difficult to control the temperature of the enzyme perfusion, and it is difficult to remove the liver after digestion.

At the same time, this technical solution is not suitable for long-clawed gerbil HSC

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