A microengineered collagen scaffold for generating a polarized crypt-villus architecture of human small intestinal epithelium

Abstract

The human small intestinal epithelium possesses a distinct crypt-villus architecture and tissue polarity in which proliferative cells reside inside crypts while differentiated cells are localized to the villi. Indirect evidence has shown that the processes of differentiation and migration are driven in part by biochemical gradients of factors that specify the polarity of these cellular compartments; however, direct evidence for gradient-driven patterning of this in vivo architecture has been hampered by limitations of the in vitro systems available. Enteroid cultures are a powerful in vitro system; nevertheless, these spheroidal structures fail to replicate the architecture and lineage compartmentalization found in vivo, and are not easily subjected to gradients of growth factors. In the current work, we report the development of a micropatterned collagen scaffold with suitable extracellular matrix and stiffness to generate an in vitro self-renewing human small intestinal epithelium that replicates key features of the in vivo small intestine: a crypt-villus architecture with appropriate cell-lineage compartmentalization and an open and accessible luminal surface. Chemical gradients applied to the crypt-villus axis promoted the creation of a stem/progenitor-cell zone and supported cell migration along the crypt-villus axis. This new approach combining microengineered scaffolds, biophysical cues and chemical gradients to control the intestinal epithelium ex vivo can serve as a physiologically relevant mimic of the human small intestinal epithelium, and is broadly applicable to model other tissues that rely on gradients for physiological function.

Advances in Hydrogels in Organoids and Organs‐on‐a‐Chip

“…These hydrogels have been regarded as promising substitutes to conventional substrate materials for cell growth, with the specific features of high biocompatibility, permeability and tunable physiochemical properties. In order to shape hydrogels into stable structures and create sophisticated microchannel networks, various microfabrication approaches, such as 3D bioprinting, photopatterning, and micromolding, can be applied in hydrogel‐based OOC construction. By combining defined hydrogels and microfabrication technologies with recent progress in the OOC area, the fabrication of functional organs can be realized, which can potentially be used as disease models or in drug screening application.…”

“…Hydrogels acted as 3D matrices or scaffolds have been incorporated into OOC to engineer human tissues with multicellular architecture and organ‐specific functions by spatial control of microenvironment cues . A variety of 3D parenchymal tissues in hydrogel‐based organs‐on‐chips have been reported, such as liver, heart, skeletal muscle, intestine, and nasal mucosa …”

Biomaterials and biosensors in intestinal organoid culture, a progress review

“…Other than Matrigel, collagen‐based hydrogels can also support the growth and polarization of intestinal epithelia. Wang et al () fabricated a collagen scaffold with micropatterned cylindrical protrusions and depressions. The intestinal epithelia seeded on the scaffold can undergo polarization and recreate crypt‐villus architecture.…”

Host‐microbe interaction in the gastrointestinal tract

“…4). In these models, the basal cell side in contact with the silicone is not accessible for studies on transport across the epithelial barrier or epithelial cell interactions with the vascular, nervous or immune systems, however this may be resolved by using micro-patterned collagen porous scaffolds with accessible luminal and basal cell sides (Wang et al, 2017).…”