Establishment and Application of Peristaltic Human Gut-Vessel Microsystem for Studying Host–Microbial Interaction

Abstract

Intestinal floras influence a lot of biological functions of the organism. Although animal model are strong tools for researches on the relationship between host and microbe, a physiologically relevant in vitro human gut model was still required. Here, a novel human gut-vessel microfluidic system was established to study the host-microbial interaction. Peristaltic motion of the cells on the chip was driven by a pneumatic pump. When intestinal epithelial cells (Caco2) were co-cultured with vascular endothelial cells (HUVECs) on the peristaltic microfluidic chip, Caco2 showed normal barrier and absorption functions after 5 days cultivation, which generally took 21 days in static Transwell models. Intestinal microvilli and glycocalyx layer were seen after 4 days cultivation, and Lactobacillus casei was successfully co-cultured for a week in the intestinal cavity. A model for intestinal damage and inflammatory responses caused by E. coli was set up on this chip, which were successfully suppressed by Lactobacillus casei or antibiotic. In summary, this human gut-vessel microfluidic system showed a good potential for investigating the host-microbial interaction and the effect and mechanism of microbiome on intestinal diseases in vitro.

Intestinal explant barrier chip: long-term intestinal absorption screening in a novel microphysiological system using tissue explants

“…28 Also in comparison to cell-based intestine on a chip devices that used FD4 to assess barrier integrity similar levels of FD4 permeability were shown in the IEBC. 58,[65][66][67][68] Taken together, these results validate the IEBC application of intestinal tissue explants as a functional barrier model.…”

“…S8a and b FITC or FITC-dextran conjugates are frequently used to assess the integrity of epithelial barriers in vitro. While many studies use large molecular weight FITC-dextran conjugates of ≥20 kDa, 16,18,21,44,[56][57][58][59][60] we chose to use a marker molecule with a much smaller molecular weight of 4 kDa. The smaller the molecule used for such measurements, the better it confirms maintained barrier integrity.…”

Exploring Effects of Chitosan Oligosaccharides on the DSS-Induced Intestinal Barrier Impairment In Vitro and In Vivo

“…Cells were seeded in 96-well plates at a density of 1 × 10 4 cells/well. After incubation, the cells were treated with different concentrations of COS, HWCOS, and NACOS (0, 10, 100, 200, 500, 1000 µg/mL) for 24 h. Then, 0.5 mg/mL MTT (M5655, Sigma-Aldrich) was added into the medium and the cells were incubated for 4 h in a 37 °C incubator as previously described [ 49 ]. Dimethyl sulfoxide (DMSO, 150 µL/well) was used to dissolve the formazan crystal.…”

“…Immunofluorescent staining of cells was performed according to the pervious study with minor revision [ 49 ]. After cultivation, the cells were treated with 4% paraformaldehyde (28908, Thermo Fisher Scientific, Waltham, MA, USA) for 30 min, and then the cells were ruptured with 0.3% Triton-X-100 at room temperature for 5 min.…”

Microfluidic Device Using Mouse Small Intestinal Tissue for the Observation of Fluidic Behavior in the Lumen

“…This is because they can reconstruct biological systems on chips and build elaborate fluidic systems with biological samples, such as cells and tissues, under microscopic observation. An in vitro biomimetic "guton-a-chip" was proposed to mimic intestinal biological functions and enable microscopic observations [16][17][18][19][20][21][22]. Those chips commonly have two microchannels in parallel, separated by a flexible and a porous membrane.…”