Skip to main content
. 2020 Jan 14;9:e50135. doi: 10.7554/eLife.50135

Figure 1. Duodenum Intestine-Chip: a microengineered model of the human duodenum.

(a) Brightfield images of human duodenal organoids (top) and human microvascular endothelial cells (bottom) acquired before their seeding into epithelial and endothelial channels of the chip, respectively. (b) Schematic representation of Duodenum Intestine-Chip, including its top view (left) and vertical section (right) showing: the epithelial (1; blue) and vascular (2; pink) cell culture microchannels populated by intestinal epithelial cells (3) and endothelial cells (4), respectively, and separated by a flexible, porous, ECM-coated PDMS membrane (5). (c) Scanning electron micrograph showing complex intestinal epithelial tissue architecture achieved by duodenal epithelium grown for 8 days on the chip (top) in the presence of constant flow of media (30 µl/hr) and cyclic membrane deformations (10% strain, 0.2 Hz). High magnification of the apical epithelial cell surface with densely packed intestinal microvilli (bottom). See Figure 1-figure supplement demonstrating the effect of mechanical forces on the cytoarchitecture of epithelial cells and the formation of intestinal microvilli (d) Composite tile scan fluorescence image 8 days post-seeding (top) showing a fully confluent monolayer of organoid-derived intestinal epithelial cells (magenta, ZO-1 staining) lining the lumen of Duodenum Intestine-Chip and interfacing with microvascular endothelium (green, VE-cadherin staining) seeded in the adjacent vascular channel. Higher magnification views of epithelial tight junctions (bottom left) stained against ZO-1 (magenta) and endothelial adherence junctions visualized by VE-cadherin (bottom right) staining. Cells nuclei are shown in gray. Scale bars, 1000 µm (top), 100 µm (bottom) (e) Apparent permeability values of Duodenum Intestine-Chips cultured in the presence of flow and stretch (30 µl/hr; 10% strain, 0.2 Hz) for up to 10 days. Papp values were calculated from the diffusion of 3 kDa Dextran from the luminal to the vascular channel. Data represent three independent experiments performed with three different chips/donor, total of three donors; Error bars indicate s.e.m.

Figure 1.

Figure 1—figure supplement 1. Flow-induced increase in primary intestinal epithelial cells height and microvilli formation.

Figure 1—figure supplement 1.

(a) Representative confocal images of x-y (top) and x-z (bottom) optical sections of duodenal organoid-derived epithelial cells cultured for 72 hr under static (Static), fluid flow (30 µl/hr; Flow) or flow and stretch (30 µl/hr; 10% strain, 0.2 Hz; Flow+Stretch) conditions and stained for apical marker villin (green) and basolateral protein E-cadherin (magenta). Nuclei were counterstained with DAPI (gray). Scale bar, 50 µm. (b) Quantitative analysis of the average cell height measured from Z-stack images as the distance between apical marker villin (green) and PDMS membrane (dotted line). Data represent the mean ± s.e.m; One-way ANOVA, ****p<0.0001. (c) Scanning electron microscopy surface images of duodenal organoid-derived epithelium cultured for 72 hr under static or flow +/- stretch conditions. Cells seeded in the top channel of Intestine-Chip were maintained with (Flow) or without (Static) medium perfusion (30 µl/hr) in both channels and 10% of mechanical stretch (0.2 Hz) (Flow+Stretch). Images were captured at the center area of the chamber. Scale bar, 5 µm. (d) Quantification of microvilli. Density of microvilli per µm2 was measured from the SEM images (100 µm2, 20 FOV) as described in the Materials and methods. Data represent the mean ± s.e.m; one-way ANOVA, ****p<0.0001.