Membrane-free culture and real-time barrier integrity assessment of perfused intestinal epithelium tubes

Abstract: In vitro models that better reflect in vivo epithelial barrier (patho‐)physiology are urgently required to better predict adverse drug effects ahead of clinical trials. Here, we describe extracellular matrix‐supported intestinal tubules, continuously-perfused in a high-throughput microfluidic system. These tubules exhibit key tissue polarization markers (ErB1, ErBb2, Ezrin, ZO-1), relevant transporters (Glut-2, MRP2), and crypt-like morphology. Leak-tight polarization of these gut-on-a-chip barrier tubules was achieved after just 4 days in culture, and high-content characterization of the tubules was made possible due to the absence of any artificial membranes in the microfluidics.
Recently, functional compound-induced toxicity was assessed by apical exposure of model compounds to these in vitro intestinal barrier tissues. Within each microfluidic system, forty leak-tight intestinal tubules were cultured in parallel, and their response to pharmacological stimuli was recorded over 125 hours of exposure. Overall, this study comprised 357 gut-on-a-chip tubes, of which 93% were confirmed to be leak tight before exposure. Kaplan-Meier plots and EC50‐time curves could be extracted from the leakage datasets, providing insight into both concentration and exposure time response.
Full compatibility with standard equipment and user‐friendly operation make this gut‐on‐a‐chip platform readily applicable in routine laboratories and high-throughput screening facilities, allowing prediction of intestinal toxicity far ahead of costly animal and clinical trials.