In-vitro model construction method for rapidly screening macromolecular immunocompetent substances

Abstract

The invention relates to an in-vitro model construction method for rapidly screening macromolecular immunocompetent substances, which comprises the following steps: preparing a qualified conditioned culture medium, and stimulating intestinal epithelial cells by using the conditioned culture medium; then simultaneously adding a sample to be detected (upper chamber) and immune perception cells RAW264.7 (lower chamber) into Caco-2 intestinal epithelium monolayer cells for co-culture and incubation; after incubation, discarding the culture medium in the upper chamber, and adding a sample to be detected and a conditioned culture medium into the upper chamber; collecting the lower room culture medium, and detecting the level of inflammatory factors. According to the method, intestinal epithelial cells and immune cells are co-cultured to simulate the environment in the intestinal tract, and a sample to be screened and a conditioned culture medium jointly act on the epithelial cells on the upper layer to simulate the intestinal injury and organism immune regulation effect of food macromolecules in the intestinal tract.

Description

technical field [0001] The invention belongs to the technical field of immune substances, in particular to an in vitro model construction method and application method for rapidly screening macromolecular immune active substances. Background technique [0002] In recent years, a large number of research results have shown that natural macromolecules such as polysaccharides and polypeptides have immunomodulatory activities. Take polysaccharides as an example. As one of the most important biological macromolecules, they are widely found in plants, animals and microorganisms. They are rich in variety and have huge reserves, and polysaccharides from different sources have different functions. For example, aloe polysaccharides promote immune cells to produce cytokines , Panax notoginseng polysaccharide activates the complement system, Indonesian curcuma polysaccharide improves macrophage phagocytosis, etc. Therefore, it is of great significance to screen macromolecular substance...

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Problems solved by technology

[0003] However, most of the existing screening models for the immune activity of macromolecules directly stimulate immune cells, and then detect the activity of immune cells and the level of inflammatory factors, etc., but it is difficult for these macromolecules to directly pass through intestinal epithelial cells to activate immune cells and regulate the biological activity of immune cells, so this screening model is not reasonable (Luo et al.,Characterization and immunological activity of polysaccharides from Ixerispolycephala 113(2018)804-812)

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