Modeling Biochemical Gradients In Vitro to Control Cell Compartmentalization in a Microengineered 3D Model of the Intestinal Epithelium

• Published in: Advanced healthcare materials Vol. 11; no. 22; pp. e2201172 - n/a
• Main Authors: Altay, Gizem, Abad‐Lázaro, Aina, Gualda, Emilio J., Folch, Jordi, Insa, Claudia, Tosi, Sébastien, Hernando‐Momblona, Xavier, Batlle, Eduard, Loza‐Álvarez, Pablo, Fernández‐Majada, Vanesa, Martinez, Elena
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01-11-2022; John Wiley and Sons Inc
• Subjects: 3D architectures; Biochemistry; biomolecular gradients; Cell differentiation; Cell Differentiation - physiology; engineering organoids; Epithelial cells; Epithelium; Finite element method; Fluorescence microscopy; Hydrogels; in silico modeling; Intestinal Mucosa - metabolism; intestinal stem cell niches; Intestine; Intestine, Small; Intestines; light sheet fluorescence microscopy; Mechanical properties; Modelling; Organoids; Organoids - metabolism; photolithography; Small intestine; Stem cells; Stroma; Three dimensional models; Villus; 3D architectures; photolithography; intestinal stem cell niches; in silico modeling; engineering organoids; biomolecular gradients; light sheet fluorescence microscopy
• ISSN: 2192-2640; 2192-2659; 2192-2659
• DOI: 10.1002/adhm.202201172

Gradients of signaling pathways within the intestinal stem cell (ISC) niche are instrumental for cellular compartmentalization and tissue function, yet how are they sensed by the epithelium is still not fully understood. Here a new in vitro model of the small intestine based on primary epithelial cells (i), apically accessible (ii), with native tissue mechanical properties and controlled mesh size (iii), 3D villus‐like architecture (iv), and precisely controlled biomolecular gradients of the ISC niche (v) is presented. Biochemical gradients are formed through hydrogel‐based scaffolds by free diffusion from a source to a sink chamber. To confirm the establishment of spatiotemporally controlled gradients, light‐sheet fluorescence microscopy and in‐silico modeling are employed. The ISC niche biochemical gradients coming from the stroma and applied along the villus axis lead to the in vivo‐like compartmentalization of the proliferative and differentiated cells, while changing the composition and concentration of the biochemical factors affects the cellular organization along the villus axis. This novel 3D in vitro intestinal model derived from organoids recapitulates both the villus‐like architecture and the gradients of ISC biochemical factors, thus opening the possibility to study in vitro the nature of such gradients and the resulting cellular response.
Biomolecular gradients are fundamental in intestinal epithelial tissue organization and function. The microengineered platform developed, with well‐controlled spatiotemporal gradients of intestinal stem cell niche factors coming from the stroma, 3D architecture and organoid‐derived cells retrieves an in vivo‐like cell compartmentalization. The system is easily incorporated in standard culture inserts where one can systematically test factors critical for intestinal tissue dynamics.