Development of a primary human Small Intestine-on-a-Chip using biopsy-derived organoids

TLDR

The study presents a method to fabricate a primary human Small Intestine‑on‑a‑Chip using epithelial cells derived from healthy intestinal biopsies. Epithelial cells were expanded as 3D organoids, dissociated, and seeded onto a porous membrane in a microfluidic chip, while parallel microchannels cultured human intestinal microvascular endothelium under flow and cyclic stretch. The chip supports villi‑like, polarized epithelium that differentiates into multiple lineages, interfaces with endothelium, and transcriptomic profiling shows it more closely resembles in vivo duodenum than organoids, enabling continuous fluid sampling for nutrient digestion, mucus secretion, barrier function, and making it a versatile tool for metabolism, nutrition, infection, drug pharmacokinetics, and personalized medicine studies.

Abstract

Abstract Here we describe a method for fabricating a primary human Small Intestine-on-a-Chip (Intestine Chip) containing epithelial cells isolated from healthy regions of intestinal biopsies. The primary epithelial cells are expanded as 3D organoids, dissociated, and cultured on a porous membrane within a microfluidic device with human intestinal microvascular endothelium cultured in a parallel microchannel under flow and cyclic deformation. In the Intestine Chip, the epithelium forms villi-like projections lined by polarized epithelial cells that undergo multi-lineage differentiation similar to that of intestinal organoids, however, these cells expose their apical surfaces to an open lumen and interface with endothelium. Transcriptomic analysis also indicates that the Intestine Chip more closely mimics whole human duodenum in vivo when compared to the duodenal organoids used to create the chips. Because fluids flowing through the lumen of the Intestine Chip can be collected continuously, sequential analysis of fluid samples can be used to quantify nutrient digestion, mucus secretion and establishment of intestinal barrier function over a period of multiple days in vitro . The Intestine Chip therefore may be useful as a research tool for applications where normal intestinal function is crucial, including studies of metabolism, nutrition, infection, and drug pharmacokinetics, as well as personalized medicine.