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

• Published in: Nature communications Vol. 8; no. 1; pp. 262 - 8
• Main Authors: Trietsch, Sebastiaan J., Naumovska, Elena, Kurek, Dorota, Setyawati, Meily C., Vormann, Marianne K., Wilschut, Karlijn J., Lanz, Henriëtte L., Nicolas, Arnaud, Ng, Chee Ping, Joore, Jos, Kustermann, Stefan, Roth, Adrian, Hankemeier, Thomas, Moisan, Annie, Vulto, Paul
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 15.08.2017; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/277; 692/4020/2199; Acetylsalicylic acid; Biochips; Caco-2 Cells; Cell culture; Cell Culture Techniques - instrumentation; Cell Culture Techniques - methods; Collagen; Epithelium; Extracellular matrix; Humanities and Social Sciences; Humans; In vitro methods and tests; Integrity; Intestinal Mucosa - chemistry; Intestinal Mucosa - metabolism; Intestine; Kinetics; Lab-On-A-Chip Devices; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Microfluidics; multidisciplinary; Permeability; Science; Science (multidisciplinary); Side effects; Stains & staining; Staurosporine; Tubes; Tubules
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-017-00259-3

In vitro models that better reflect in vivo epithelial barrier (patho-)physiology are urgently required to predict adverse drug effects. Here we introduce extracellular matrix-supported intestinal tubules in perfused microfluidic devices, exhibiting tissue polarization and transporter expression. Forty leak-tight tubules are cultured in parallel on a single plate and their response to pharmacological stimuli is recorded over 125 h using automated imaging techniques. A study comprising 357 gut tubes is performed, of which 93% are leak tight before exposure. EC 50 -time curves could be extracted that provide insight into both concentration and exposure time response. Full compatibility with standard equipment and user-friendly operation make this Organ-on-a-Chip platform readily applicable in routine laboratories. Efforts to determine the effects of drugs on epithelial barriers could benefit from better in vitro models. Here the authors develop a microfluidic device supporting the growth and function of extracellular matrix-supported intestinal tubules, and evaluate the effect of staurosporine and acetylsalicylic acid on barrier integrity.