Immunomodulatory effects of dietary supplements on IPEC-J2 cells

Prebiotics are used by the gut microbiota creating metabolites that exert a positive effect on the host (postbiotics). Prebiotics are available on the market for human and animal consumption and may induce differential effects on the host's health and metabolism. They can be tested by in vitro methods to evaluate their capacity to exert a beneficial effect on gut inflammation and barrier by using in vitro assays and cell cultures. IPEC-J2 cells are intestinal epithelial cells non-transformed obtained from the jejunal segment from a newborn and un-suckled piglet. Cells are grown in flat bottom flasks/plates at 37°C and 5% CO2 humidified atmosphere in complete DMEM/F-12 supplemented with 1% penicilin-streptomicyn, 5% fetal bovine serum, Lglutamine, EGF, Isulin, transferrin and selenium. Screening of cytotoxicity of the products of interest by MTT test in a concentration series of the product of interest to establish the dose. Incubation for 24 hours with the chosen concentration of product and parallel LPS challenge. Collection of supernatants and cells for the measurement of the immune mediators released in the medium by the cells, such as cytokines. Collection of cells for RNA extraction and evaluation of gene expression changes induced in the cells by qPCR methods. In parallel, cells can be cultured in snapwell for studying barrier integrity properties by measuring permeability changes and transepithelial resistance in Ussing chambers.