(1004-A) A high-throughput gut-on-a-chip model integrating adult stem cell-derived organoids for disease modeling and drug discovery

Abstract: Adult stem cell derived organoids are a rapidly rising technology with the potential to revolutionize drug discovery. Standard organoid culture protocols usually rely on embedding stem cells in an extracellular matrix which enables their growth and differentiation as polarized cystic structures. While this culture protocol yields organoids with biological similarity with the original organ, it concurrently presents several limitations, such as inherent organoid heterogeneity, difficult experimental manipulation, limited apical access and lack of perfusion. Microfluidic techniques are increasingly recognized as an important toolbox to add physiological parameters to cell culture while enabling precise patterning and architecture. Hence, integrating organoid technology with microfluidics offers the possibility to rely on the self-assembly properties of organoids, while simultaneously inducing a tailored architecture allowing for apical access, perfusion, automated procedures and ease of experimental handling. Yet, standardized microfluidic organoid models are still in their infancy and their use for high throughput screening and drug development remains unexplored.
We established a high throughput capable human gut-on-a-chip model composed of intestinal organoids-derived epithelial cells patterned inside of the microfluidic channel of a MIMETAS OrganoPlate®. Cells introduced into the system adhere to an ECM boundary and rapidly populate the entirety of the microfluidic channel forming tubular structures. The tubular architecture allows for apical access, as well as enables gravity driven fluid flow. We observed rapid cell polarization as indicated by cellular localization of marker expression as well as directional transporter activity. This revealed the presence of an epithelial apical side oriented toward the tubule lumen opposed to the basolateral side facing the ECM substrate. We confirmed the expression of major intestinal markers such as MUC2, ZO-1, villin, or lysozyme. We also observed functional enzymatic activity associated with drug metabolism. Tight junction formation, as well as transepithelial electrical resistance (TEER) showed high barrier function with robust reproducibility. Moreover, we showed that several readouts such as immunostaining and TEER could be used to evaluate the intestinal response to the gut tubules after exposure to toxic compounds and inflammatory triggers.
Through the combination of adult human stem cell-derived intestinal organoids and a microfluidic platform compatible with automation, we present a powerful system to study physiology and disease mechanisms. Such a system could be used to perform drug screenings in a patient-specific gut model, ultimately improving our ability to develop tailored therapeutic interventions.