Method for separating liver cells and hepatic stellate cells in animals infected with echinococcus multilocularis

Abstract

The invention relates to a method for separating liver cells from hepatic stellate cells in animals infected with echinococcus multilocularis. The method for separating liver cells from hepatic stellate cells in animals infected with echinococcus multilocularis comprises the following steps of: (1) in-situ perfusion: performing in-situ perfusion in the liver of animals infected with echinococcus multilocularis by sequentially using a pre-perfusion liquid, a post-perfusion liquid 1 and a post-perfusion liquid 2 through a portal vein, and taking off the liver to prepare an intrahepatic cell suspension; and (2) gradient centrifugation: centrifuging the intrahepatic cell suspension to obtain a supernatant 1 and a precipitate 1 at the lower layer, centrifugally washing the precipitate 1 with D-Hanks solution to obtain hepatocytes, centrifuging the supernatant 1, collecting a precipitate 2 at the lower layer, sequentially adding GBSS-B, Nycodenz separation solution and GBSS-B into the precipitate 2, performing centrifuging, collecting white annular layer, centrifugally centrifugal washing with GBSS-B, and discarding supernatant to obtain the hepatic stellate cells. The method for separating liver cells from hepatic stellate cells in animals infected with echinococcus multilocularis can extract and separate the liver cells and the hepatic stellate cells in the animals infected with echinococcus multilocularis at the same time.

Description

technical field [0001] The invention belongs to the technical field of liver cell separation, and in particular relates to a method for separating hepatic cells and hepatic stellate cells in alveolar coccidia infected animals. Background technique [0002] Liver fibrosis is a complex disease involving multiple cell types and cytokines in the liver. The liver is composed of hepatic parenchymal cells and hepatic non-parenchymal cells, such as hepatocytes, hepatic stellate cells, Kupffer cells, and endothelial cells. Hepatic cell apoptosis is rare in normal liver tissue. With the occurrence and development of liver fibrosis, the apoptosis of hepatic cells will change significantly. . There is a close causal relationship between hepatocyte apoptosis, HSC activation and liver fibrosis. It has been reported in the literature that under the action of some injury factors, apoptosis of hepatocytes becomes the first response of cells to toxins, and these apoptotic hepatocytes relea...

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Problems solved by technology

[0005] However, using the above-mentioned existing methods to isolate hepatocytes or hepatic stellate cells in the animal model infected with alveolar coccidia, there is not only the technical problem of not being able to obtain hepatocytes and hepatic stellate cells at the same time, but also obtaining the same number of viable cells. There are also disadvantages of poor separation effect of hepatic cells or hepatic stellate cells, low survival rate and purity

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