The human gut microbiota constitutes the most bountiful and divergent community of organisms compared to the other areas of the body . Growing attention is devoted to the bacterial equilibrium and constitution in human intestines, being highly plastic over time [2,3]. Bacteria present a significant part in response to immunotherapy in cancer . Meanwhile, predominant cell cultures and animal models encounter substantial limitations .
3D-multi-compartment microfluidic cultures may overcome met obstacles, mimicking elaborate multicellular architectures and niches, while maintaining high control. Among the numerous gut-on-chip designs, the differentiation between 2D and 3D cellular microenvironments is possible, either mechanically stimulated or not. Mechanobiological actuation and 3D niche recapitulation proved integral in maturing and translating models from in vivo to in vitro [6,7,8].
Here, we present the intestine-on-chip PDMS-based device with endothelium captured in 3D under mechanobiological stimulation. As a proof-of-concept, the device integrates actuation on a three-cell types coculture, which includes human micro-endothelial cells and two human intestinal epithelial cell lines endowed with different adsorbing and secretory properties, respectively. A collagen-based extracellular-matrix compartment connects the intestinal and the vascular spaces. Multi-parametric assessment of cell viability and function at different time points describes the influence of mechanobiological stimuli on the maturation of the gut endothelium. Transcriptional profiling of the epithelial cells and functional characterization of different primary immune cells illustrate the suitability of the system to dissect complex interactions between components of the tumour microenvironment.
The biomechanically stimulated 3D intestine-on-chip provides an elegant platform to study how microorganisms inflect the crosstalk between epithelial and endothelial compartments in the gut and portray a relevant alternative for preclinical studies. The acquired model allows dissecting the trans-endothelial migration of immune cells in health and disease.
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