Developing a Luminal Perfusion System for Human Intestinal Organoids

Abstract

Human intestinal organoids (HIOs) are 3-dimensional aggregates of cells that can replicate the structure and function of the human gastrointestinal system. HIOs generally resemble fetal tissue, so efforts have been made to mature them such that they replicate adult gastrointestinal physiology. Previous studies have shown HIO maturation following an injection of E. coli into the HIO lumen but did not attempt to regulate E. coli populations thereafter. This thesis presents a perfusion system which has the potential to modulate a luminal bacteria population through fluidic transfer. The system consists of a double-barrel glass capillary connected to a pressure-based pump, which allows for high temporal control of fluid flow. The system has been shown to regulate the concentration of diffusible dye molecules in the lumen and partially regulate E. coli distributed heterogeneously in the lumen. If volumetric flow control and incubator compatibility are implemented, the new proposed system may enable long-term study of HIO-E. coli interaction.