Intestinal organoids are a key innovative technology (ST2) to replace animal testing and measure complex traits in animal breeding and nutrition. Organoids are three-dimensional (3D) structures that represent the organ from which they originate. Organoids can be used to study complex traits such as feed efficiency and gut health. For example, organoids provide an in vitro research platform for comparative testing of nutrient or drug absorption between individuals. The use of organoids in animal breeding, nutrition and health will have a major impact on improving priorities in the Topsector missions “circular agriculture” (A) and “Appreciated, healthy and safe food” (D). This project follows the successful launch of the PPP project (LWV20.25), where we demonstrated that we can generate 3D and 2D intestinal organoids from pigs. Furthermore, the 2D Transwell model using pig intestinal organoids were shown to be functional, as demonstrated by nutrient absorption and barrier function testing. However, follow-up research is needed to test different functional ingredients and utilizing variation between individuals. To improve the in vivo emulation of these models, a next level of complexity is required. Interaction of epithelial cells with immune cells are not regarded in the current organoid models and the biological signaling pathways are still largely unknown, which are important components for understanding gut health. Co-culturing of pig intestinal organoids with immune cells and gene editing of organoids will give insight in the interaction between epithelial and immune cells. Therefore, the main goals of the new PPP are to validate (1) the already developed organoid based Transwell system for nutrition absorption and test different functional ingredients using organoids derived from phenotypically diverse pigs; develop (2) co-culturing of pig intestinal organoids and immune cells, and (3) gene editing in organoids to get understanding of the biological signaling pathways to
improve the application of the in vitro organoid system in animal breeding and nutrition research. This project consists of three work packages (WP). In WP1, the developed Transwell model will be validated to test nutrient absorption and investigate functional capacity of organoids derived from phenotypically diverse pigs. In WP2, we will investigate the possibility of developing a pig intestinal organoid immune cell co-culture model system. In WP3, we will develop a model for the application of gene editing using intestinal organoids. Results of this project will lead to a more robust and versatile in vitro system that will allow us to measure complex traits and test the bio-functionality of various dietary components, including nutrients. One of the advantages of such a model is that it retains the genetic information of the individual pig, which can help researchers plan in vitro studies using organoids from different pigs that represent the variation present in the pig population. We anticipate that the use of such a near-physiological in vitro system will result in fewer animals needed for experiments and will contribute to animal breeding and nutrition research.
|Effective start/end date||1/01/23 → 1/01/27|