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.
Highlights
The small intestine is the major site for digestion, drug and nutrient absorption, interaction with commensal microbiome, and development of mucosal immunity, as well as a primary site for many diseases, such as bacterial, viral and parasitic infections and inflammatory bowel disease
We report here the development of a bioengineered in vitro model of the duodenum portion of the human small intestine by combining microfluidic Organ Chip technology with organoid-based methods for culture of primary epithelial stem cells from duodenal biopsies
The Intestine Chip recapitulates important structural features and functions of the native duodenum, including formation of elongated villi-like structures lined by a polarized epithelium expressing markers of enterocytes, goblet cells, enteroendocrine cells and Paneth cells, as well as basal proliferative cells, strong barrier function, brush border digestive enzyme activity, and continuous accessibility to fluids flowing through the apical lumen
Summary
The small intestine is the major site for digestion, drug and nutrient absorption, interaction with commensal microbiome, and development of mucosal immunity, as well as a primary site for many diseases, such as bacterial, viral and parasitic infections and inflammatory bowel disease. These chips recapitulate normal tissue-tissue interfaces and mimic the complex physical and biochemical microenvironment of living human organs[9,10,11,12,13,14,15,16,17,18,19,20,21,22,23] This technology has been previously applied to develop human Gut Chips that emulate many features of human intestinal structure and function, these studies utilized established human intestinal cell lines, such as Caco-2 or HT-29 cells[19,20,21,24], which were originally isolated from tumor samples and they harbor multiple gene mutations. The transcriptome of the primary Intestine Chip more closely resembles that of adult human duodenum in vivo than the organoids that were used to plate the chips or other currently available intestinal cell culture models, including our previous Caco-2 Gut Chip, especially with regard to expression of genes relating to digestion, response to nutrients, cell proliferation, and host defense response to infection
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