Streamlined production of micropatterned scaffolds from 3D printed templates for gut in vitro models

Intestinal cancers represent exceptionally lethal malignancies with limited treatment options. To understand their oncogenesis, support the development of more effective therapies and perform ex vivo drug testing, a great effort has been invested in obtaining tissue-like miniguts-on-chip. Current in vitro models, such as intestinal organoids, have limitations including restricted luminal access, short life-span and inadequate representation of the tumour microenvironment (TME). By combining template micropatterning with collagen-based scaffolds, this work aims to create in vitro 3D coltures recapitulating the intestinal epithelium. To this aim, we engineered by micropatterning hydrogel scaffolds with crypts and villi structures. The process involves the fabrication of a master template using 3D printing, the generation of an elastomeric replica, followed by the production of crosslinked collagen-based scaffolds. These scaffolds are suitable for cell culture, as shown by the functional growth of CaCo-2 polarised epithelia, and display architectural resemblance to in vivo intestinal tissues. By integrating these scaffolds into a bicompartmental culture system, known as the True Tissue on Platform (TToP), we want to use this 3D coltures to model the human intestinal tissue and assess the efficacy of anticancer drugs.