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Imprint
Generation and characterization of 3D-organoids from bovine colon material to investigate the interactions of zoonotic bacteria with the host mucosa. Subproject: Development of 'hangig drops' organoid cores from intestinal fibroblasts and edited primary epithelial cells (3D-COR)
Project
Project code: 01KI1732B
Contract period: 01.01.2018
- 29.02.2020
Budget: 34,800 Euro
Purpose of research: Experimental development
Shiga toxin-producing Escherichia coli (STEC) are globally distributed and among the most frequently transmitted foodborne pathogens. When infecting humans, STEC can cause a wide variety of different diseases. Ruminants, in particular cattle, represent the most important natural reservoir for STEC. Some strains are highly zoonotic, as evidenced by outbreaks that can be traced epidemiologically to cattle herds. However, they are often isolated only sporadically from humans and cattle. In contrast, other strains can persist in cattle or cattle herds for long periods of time. They might serve as continuous source of virulence factors in the generation of novel outbreak strains. Persisting STEC strains have to colonize a specific niche in the host for an extended time span. Here, attachment to the intestinal mucosa as well as multiple interactions between bacterial and host cells both make important contributions to colonization. Immortalized cell lines or 2D-cell culture models are simplified model systems that cannot reproduce many details of the in vivo situation. To bridge the gap between cell lines and animal research, for developing pre-clinical animal models to the point of replacing animal experiments entirely, requires more sophisticated cellular in vitro models. Stem cell-based intestinal “mini-organs” (3D-organoids) are being used extensively to investigate regulatory and pathological mechanisms of the intestinal epithelium. Such 3D-organoids have not yet been derived from cattle intestinal tissue. Therefore, the goal of this pilot project is to establish a three-dimensional cell culture system with cells isolated from the colon of calves. These 3D-organoids will be used to characterize the colonization and the interactions of persisting and non-persisting STEC strains with the cattle reservoir hosts. They will also serve as model system in the identification of genes from both bacteria and host that contribute to persistent colonization.
Section overview
Subjects
- Animal health
- Biotechnology
