Project description:Entamoeba histolytica, the agent of amebiasis, colonizes the human colon and can invade the lining of the colon to disseminate in the deep layers of the intestine. Amebiasis mainly affects poor people in developing countries, where the barriers between human feces and food or water are inadequate. Humans are the only reservoir of E. histolytica and are the sole target organism of the development of the disease, which limits our knowledge of the crosstalk between the colon and the parasite, especially during the acute phase of amebiasis. In the present work, we constructed a three-Dimensional (3D)-intestinal model capable of reproducing important features of the human intestine. Using this model and leading-edge technologies, including tissue and cell imaging, transcriptomics, and proteomics, as well as ELISA-based immune response inspection, the early stages of amebic infection have been studied. The data obtained highlight the importance of several virulence markers already shown in patients or experimental models, but also underscore the involvement of other factors that appear to be key regulators of gene expression or important in the secretome of the infected tissue. In addition, we characterized the cellular stress responses against amebiasis and novel regulatory mechanisms utilized by this parasite to modulate the immune response and survive within the human intestine.
Project description:Investigations of human parasitic diseases depend on the availability of appropriate in vivo animal models and ex vivo experimental systems, and are particularly difficult for pathogens whose exclusive natural hosts are humans, as for Entamoeba histolytica, the protozoan parasite responsible for amoebiasis. We elaborated a human hepatic in vitro model consisting of cultured liver sinusoidal endothelial cells and hepatocytes in a 3D collagen-I environment. We have characterized the model's barrier function, its chemical and structural complexity and examined E. histolytica invasion. This study deals with secretome of hepatic model under various invasion conditions.
Project description:Here we describe a method for fabricating a primary human Small Intestine-on-a-Chip (Intestine Chip) containing epithelial cells isolated from healthy regions of intestinal biopsies. The primary epithelial cells are expanded as 3D organoids, dissociated, and cultured on a porous membrane within a microfluidic device with human intestinal microvascular endothelium cultured in a parallel microchannel under flow and cyclic deformation. In the Intestine Chip, the epithelium forms villi-like projections lined by polarized epithelial cells that undergo multi-lineage differentiation similar to that of intestinal organoids, however, these cells expose their apical surfaces to an open lumen and interface with endothelium. Transcriptomic analysis also indicates that the Intestine Chip more closely mimics whole human duodenum in vivo when compared to the duodenal organoids used to create the chips. Because fluids flowing through the lumen of the Intestine Chip can be collected continuously, sequential analysis of fluid samples can be used to quantify nutrient digestion, mucus secretion and establishment of intestinal barrier function over a period of multiple days in vitro. The Intestine Chip therefore may be useful as a research tool for applications where normal intestinal function is crucial, including studies of metabolism, nutrition, infection, and drug pharmacokinetics, as well as personalized medicine.
Project description:Applicability of in vitro (human Caco-2 cells) and ex vivo intestine models (rat precision cut intestine slices and the pig in-situ small intestinal segment perfusion (SISP) technique) to study the effect of food compounds. In vitro digested yellow (YOd) and white onion extracts (WOd) were used as model food compounds and transcriptomics was applied to obtain more insight into which extent mode of actions depend on the model.
Project description:Rat small intestine precision cut slices were exposed for 6 hours to in vitro digested yellow (YOd) and white onion extracts (WOd) that was followed by transcriptomics analysis. The digestion was performed to mimic the digestion that in vivo takes place in the stomach and small intestine. The transcriptomics response of the rat small intestine precision cut slices was compared to that of human Caco-2 cells and the pig in-situ small intestinal segment perfusion. The microarray data for the human Caco-2 cells (GSE83893) and the pig in-situ small intestinal segment perfusion (GSE83908) have been submitted separately from the current data on rat intestine. The goal was to obtain more insight into to which extent mode of actions depend on the experimental model. A main outcome was that each of the three models pointed to the same mode of action: induction of oxidative stress and particularly the Keap1-Nrf2 pathway.
Project description:The goal of this study was to compare RNA expression of the A) CEA gene obtained from i) colon intestine organoids cultured in conventional plastic-adherent Matrigel drop overlaid with growth medium; ii) Colon Intestine-Chips derived from organoids in the presence of constant flow and stretch; iii) human colon intestine tissue and to verify whether Colon Intestine-Chip faithfully recapitulates human duodenum tissue and to better understand how much it differs from the organoids from which it’s derived, and B) FLOR1 gene obtained from i) human alveolar epithelial cells in T-25 Flasks, ii) human Alveolus Lung-Chip on day 5 and day 10 of the culture.