Biological processes induced by ZnO, Amoxicillin, and Rye in cultured Intestinal Porcine Epithelial Cells (IPEC-J2). Sus scrofa

The objective of this study is to investigate to what extent gene expression data of dietary interventions generated in IPEC-J2 in vitro model overlap with in vivo data. Gene expression was recorded in IPEC-J2 cells upon exposure to three different dietary interventions commonly used in livestock. In a further step, we compared the results with mucosal gene expression responses, as measured in animals exposed to the same compounds via the diet. As compounds we used zinc oxide, rye and the antibiotic amoxicillin. The GEO accession numbers of the in vivo datasets are provided in the paper "Enrichment of in vivo determined transcription data from dietary intervention studies with in vitro data provides improved insight into gene regulation mechanisms" (submitted to "Genes and Nutrition"). Overall design: IPEC-J2 cells were grown for 7 days at 37 ºC and 5% CO2 using 1:1 DMEM/Ham’s F10 1:1 medium supplemented with 5% FCS without antibiotics. For all tests, confluent monolayers were washed twice with medium without FCS (hereafter denoted as medium) and incubated for 1 hour with this medium. Hereafter, the medium was discarded and an additive dissolved/suspended in medium was added. Concentrations of 2.5% w/v Amoxicillin (brand name Octacillin), 0.03125% w/v of ZnO, and of a 3-fold diluted suspension of the diets containing 0%, 5% or 10% w/v Rye in culture medium were used for incubation of IPEC-J2 monolayers for a period of 2 and 6h. All incubations were tested in duplicate and for each type of additive duplicate control wells containing no additive (only culture medium) were incubated for 2 and 6h. After incubation total RNA was extracted. Each duplicate incubation was hybridized separately on a microarray patch. IPEC-J2 enterocyte cell line derived from the jejunum of piglets, host-feed interaction