Screening out irrelevant cell-based models of disease

The common and persistent failures to translate promising preclinical drug candidates into clinical success highlight the limited effectiveness of disease models currently used in drug discovery. An apparent reluctance to explore and adopt alternative cell-and tissue-based model systems, coupled with a detachment from clinical practice during assay validation, contributes to ineffective translational research. To help address these issues and stimulate debate, here we propose a set of principles to facilitate the definition and development of disease-relevant assays, and we discuss new opportunities for exploiting the latest advances in cell-based assay technologies in drug discovery, including induced pluripotent stem cells, three-dimensional (3D) co-culture and organ-on-a-chip systems, complemented by advances in single-cell imaging and gene editing technologies. Funding to support precompetitive, multidisciplinary collaborations to develop novel preclinical models and cell-based screening technologies could have a key role in improving their clinical relevance, and ultimately increase clinical success rates

Developing an advanced gut on chip model enabling the study of epithelial cell/fibroblast interactions

Organoids are widely used as a model system to study gut pathophysiology; however, they fail to fully reproduce the complex, multi-component structure of the intestinal wall. We present here a new gut on chip model that allows the co-culture of primary epithelial and stromal cells. The device has the topography and dimensions of the mouse gut and is based on a 3D collagen I scaffold. The scaffold is coated with a thin layer of laminin to mimic the basement membrane. To maintain the scaffold structure while preserving its cytocompatibility, the collagen scaffold was rigidified by threose-based post-polymerization treatment. This treatment being cytocompatible enabled the incorporation of primary intestinal fibroblasts inside the scaffold, reproducing the gut stromal compartment. We observed that mouse organoids, when deposited into crypts, opened up and epithelialized the scaffold, generating a polarized epithelial monolayer. Proper segregation of dividing and differentiated cells along the crypt-villus axis was achieved under these conditions. Finally, we show that the application of fluid shear stress allows the long-term culture of this intestinal epithelium. Our device represents a new biomimetic tool that captures key features of the gut complexity and could be used to study gut pathophysiology

Capillary pinning assisted patterning of cell-laden hydrogel microarrays in microchips

We present a capillary pinning technique that gives complete control on the local patterning of hydrogel structures in closed microchips. The technique relies on selective trapping of liquids at predefined locations in a microchip using capillary barriers. In selective patterning, the abrupt expansion in the cross-sectional geometry of a microchannel at capillary barriers results in a confined advancement of the liquid–air meniscus. This protocol describes a detailed procedure to design and fabricate microarrays of different hydrogel types, fabricated with photopolymerization or thermogelation. The process can be subdivided into two parts. First, a PDMS microchip containing microfeatures with customized patterns is fabricated. Second, the microchip is filled with a hydrogel precursor to be cross-linked by either photopolymerization or thermogelation. The production of the microchip takes approximately 2 days, depending on the substrate selection. Preparation of the hydrogel solutions takes 1–2 h, whereas the patterning and reaction to cross-link the hydrogels is completed in a few minutes