title: Development of an Advanced Primary Human In Vitro Model of the Small Intestine.
abstract: Intestinal in vitro models are valuable tools in drug discovery and infection research. Despite several advantages, the standard cell line-based Transwell(®) models based for example on colonic epithelial Caco-2 cells, lack the cellular complexity and transport activity associated with native small intestinal tissue. An additional experimental set-back arises from the most commonly used synthetic membranes, on which the cells are routinely cultured. These can lead to an additional barrier activity during in vitro testing. To overcome these limitations, we developed an alternative primary human small intestinal tissue model. This novel approach combines previously established gut organoid technology with a natural extracellular matrix (ECM) based on porcine small intestinal scaffold (SIS). Intestinal crypts from healthy human small intestine were expanded as gut organoids and seeded as single cells on SIS in a standardized Transwell-like setting. After only 7 days on the ECM scaffold, the primary cells formed an epithelial barrier while a subpopulation differentiated into intestinal specific cell types such as mucus-producing goblet cells or hormone-secreting enteroendocrine cells. Furthermore, we tested the influence of subepithelial fibroblasts and dynamic culture conditions on epithelial barrier function. The barrier integrity was stabilized by coculture in the presence of gut-derived fibroblasts. Compared to static or dynamic culture on an orbital shaker, dynamic culture in a defined perfusion bioreactor had an additional significant impact on epithelial cell differentiation, indicated by high prismatic cell morphology and upregulation of CYP3A4 enzyme and Mdr1 transporter activity. In summary, more physiological tissue models as presented in our study might be useful tools in preclinical research and development.
authors.affiliation: 1 Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg , Würzburg, Germany .
authors.forename: Matthias
authors.initials: M
authors.lastname: Schweinlin
authors.affiliation: 1 Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg , Würzburg, Germany .
authors.forename: Sabine
authors.initials: S
authors.lastname: Wilhelm
authors.affiliation: 1 Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg , Würzburg, Germany .
authors.forename: Ivo
authors.initials: I
authors.lastname: Schwedhelm
authors.affiliation: 1 Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg , Würzburg, Germany .
authors.forename: Jan
authors.initials: J
authors.lastname: Hansmann
authors.affiliation: 2 Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), Saarland University , Saarbrücken, Germany .
authors.forename: Rene
authors.initials: R
authors.lastname: Rietscher
authors.affiliation: 3 Department of General, Visceral, Vascular and Pediatric Surgery, University Hospital of Würzburg , Würzburg, Germany .
authors.forename: Christian
authors.initials: C
authors.lastname: Jurowich
authors.affiliation: 1 Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg , Würzburg, Germany .|4 Translational Center Würzburg "Regenerative Therapies for Oncology and Musculoskeletal Diseases" (TZKME), Würzburg Branch of the Fraunhofer Institute Interfacial Engineering and Biotechnology (IGB) , Würzburg, Germany .
authors.forename: Heike
authors.initials: H
authors.lastname: Walles
authors.affiliation: 1 Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg , Würzburg, Germany .|4 Translational Center Würzburg "Regenerative Therapies for Oncology and Musculoskeletal Diseases" (TZKME), Würzburg Branch of the Fraunhofer Institute Interfacial Engineering and Biotechnology (IGB) , Würzburg, Germany .
authors.forename: Marco
authors.initials: M
authors.lastname: Metzger
mesh_terms: D001692:Biological Transport; D002454:Cell Differentiation; D018920:Coculture Techniques; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000166:cytology*; D007421:Intestine, Small / Q000166:cytology*; D008954:Models, Biological