title: 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.
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Yuli
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Dulan B
authors.initials: DB
authors.lastname: Gunasekara
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Mark I
authors.initials: MI
authors.lastname: Reed
authors.affiliation: Department of Biomedical Engineering, University of North Carolina, Chapel Hill, NC, 27599, USA; North Carolina State University, Raleigh, NC, 27607, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: DiSalvo
authors.affiliation: Department of Genetics, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Bultman
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Sims
authors.affiliation: Department of Biomedical Engineering, University of North Carolina, Chapel Hill, NC, 27599, USA; North Carolina State University, Raleigh, NC, 27607, USA.
authors.forename: Scott T
authors.initials: ST
authors.lastname: Magness
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599, USA; Department of Biomedical Engineering, University of North Carolina, Chapel Hill, NC, 27599, USA; North Carolina State University, Raleigh, NC, 27607, USA. Electronic address: nlallbri@unc.edu.
authors.forename: Nancy L
authors.initials: NL
authors.lastname: Allbritton
mesh_terms: D000818:Animals; D002451:Cell Compartmentation; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000187:drug effects / Q000502:physiology*; D007421:Intestine, Small / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D009940:Organoids / Q000187:drug effects; D016062:Porosity; D051381:Rats; D046509:Tissue Culture Techniques; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D003094:Collagen
keywords: Crypt; Intestine; Microfabrication; Scaffold; Stem cell; Villus