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

Nature communications (2017-08-16)
Sebastiaan J Trietsch, Elena Naumovska, Dorota Kurek, Meily C Setyawati, Marianne K Vormann, Karlijn J Wilschut, Henriëtte L Lanz, Arnaud Nicolas, Chee Ping Ng, Jos Joore, Stefan Kustermann, Adrian Roth, Thomas Hankemeier, Annie Moisan, Paul Vulto
ABSTRACT

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. EC50-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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-Tubulin, Acetylated antibody produced in mouse, clone 6-11B-1, ascites fluid
Sigma-Aldrich
Fetal Bovine Serum, USA origin, Heat Inactivated, sterile-filtered, suitable for cell culture, suitable for insect cell culture, suitable for hybridoma
Sigma-Aldrich
Acetylsalicylic acid, ≥99.0%