A healthy, balanced diet has a fundamental role in preventing a large range of chronic diseases and contributes to prolong life quality with obvious benefits for the individual as well as for the society. Aquaculture production plays a substantial role in this perspective because fish is an important source of well-balanced proteins and important nutrients such as marine-derived omega-3 fatty acids. However, its sustainability generates concerns as farmed fish diet is largely based on fishmeal and fish oil. Consumer and environmental groups demand a continued move towards alternative feeds. Objective of this project is to develop a next generation 3D culture platform that accurately mimics the complex functions of the intestinal mucosa. Its purpose is to make available a technology for predicting the health and nutritional value of innovative components of aquafeeds. Current methods are lengthy, expensive and requires the use of large number of animals. Furthermore, they do not provide the knowledge of the cellular and molecular mechanisms determining the final effect of each meal on the fish. This lack of mechanistic knowledge severely limits our capacity to understand and predict the biological value of the single raw material and of their different combinations. We propose to develop new ad hoc biomaterials to create a 3D scaffold where to grow and differentiate a complete population of intestinal epithelial cells. Combining state of the art notions on fish nutrition will lead to a fully functional prototype of artificial intestine (Fish-AI) that will enable the feed industry to predict accurately and efficiently the health and nutritional value of alternative feed sources substantially improving European aquaculture sustainability and competitiveness. The project fosters cross-fertilisation and synergy among nutrition physiology, bioengineering, cell and stem cell biology to develop innovative technologies for a sustainable livestock production.