Co-culture system for liver cells and Kupffer cells and application thereof

Abstract

The invention discloses a co-culture system for liver cells and Kupffer cells and application thereof. The co-culture system is characterized in that in a Millicell double-layer culture room, 1-100 ng / mL LPS (lipopolysaccharides) is added into a culture medium, the Kuffer cells are cultured in the upper layer of the Millicell double-layer culture room, and the liver cells are cultured in the lower layer of the Millicell double-layer culture room; and half of the fresh culture medium is replaced every other day so as to keep the concentration of the LPS. The co-culture system for liver cells and Kupffer cells provided by the invention has the advantages that the Kupffer cells continuously secrete inflammatory cell factors (such as TNF-alpha) in vitro after being stimulated by the LPS; the liver cells can generate a great number of enzymes, such as AST (aspartate aminotransferase), ALT (alanine transaminase) and GGT (gamma-glutamyl transpeptidase) in the environment so as to stimulate the liver injury based on immunoreaction caused by the immune / inflammation response after the liver tissue undergoes virus infection in vivo; and moreover, the co-culture system is used for screening medicaments resisting immune liver injury according to the influence of the medicaments on the secretion amount of immune factors and the enzymatic activity.

Description

technical field [0001] The invention belongs to the technical field of in vitro cell culture, and particularly relates to a hepatocyte-Kupffer cell co-cultivation system, which is used as an immune liver injury model and applied to the screening of anti-immune liver injury drugs. Background technique [0002] Immune liver injury is usually caused by biological factors, and the common cause is viral infection, such as hepatitis B virus (HBV), hepatitis C virus (HCV) and other viral infections clinically. A large number of inflammatory cells infiltrate and generate an immune / inflammatory response, leading to immune response-based liver injury. After the virus invades the liver cells, it can be integrated with the DNA of the liver cells. When the immune cells or antiviral drugs kill the virus, they will also clear the infected liver cells, resulting in immune liver damage. Long-term repeated liver injury can lead to scar-like hyperplasia, liver fibrosis, cirrhosis, liver ascit...

AI Technical Summary

Problems solved by technology

[0010] Although the overall animal model is relatively reliable, it takes a long time. It usually takes 1 to 2 months from the beginning of modeling to the end of the experiment; in vitro liver cell immune injury models use a single type of cell, which is difficult to simulate in vivo complex physiological and pathological conditions

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