The Progress of Intestinal Epithelial Models from Cell Lines to Gut-On-Chip (original) (raw)
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Biomimetic human Gut-on-a-Chip is a microengineered in vitro model of human intestine that reconstitutes the intestinal microenvironment inhabited by viable gut microbiome under the precise control of mechanical deformation and fluid flow mimicking peristaltic bowel movement. By harnessing the versatile modularity of the gut-on-a-chip, key causative factors of inflammatory bowel disease (IBD) such as epithelial cells, gut microbiome, immune components, and microenvironmental cues can be independently, and collectively, manipulated for dissecting the disease mechanism. Innovations using the gut-on-a-chip may revolutionize the future treatment of IBD by validating the efficacy and safety of new IBD treatments in "Your gut-on-a-chip" that employs multiple types of host and microbial cells isolated from individual IBD patients. Ultimately gut-on-a-chip microphysiological system aims to contribute to the Precision Medicine Initiative and customized healthcare for the rescue of ...
Microfluidic Organ-on-a-Chip Models of Human Intestine
Cellular and molecular gastroenterology and hepatology, 2018
Microfluidic organ-on-a-chip models of human intestine have been developed and used to study intestinal physiology and pathophysiology. In this article, we review this field and describe how microfluidic Intestine Chips offer new capabilities not possible with conventional culture systems or organoid cultures, including the ability to analyze contributions of individual cellular, chemical, and physical control parameters one-at-a-time; to coculture human intestinal cells with commensal microbiome for extended times; and to create human-relevant disease models. We also discuss potential future applications of human Intestine Chips, including how they might be used for drug development and personalized medicine.
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Open Research Europe
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Development of a primary human Small Intestine-on-a-Chip using biopsy-derived organoids OPEN
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. 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. Because the lack of human-relevant responses has rendered many animal models unsuitable to study causal factors and treatment strategies for human intestinal infections and disorders 1 , three-dimensional (3D) human tissue surrogates, such as intestinal organoids (also known as enteroids) have emerged as promising alternatives. These spheroidal ex vivo tissue cultures include Lgr5 + intestinal stem cells 2 and are grown embedded within a complex extracellular matrix (ECM) gel (Matrigel) with Wnt3a, epidermal growth factor (EGF), Noggin and R-spondin 1 (collectively, WENR) to support their indefinite propagation 3,4. Organoids faithfully recapitulate the cellular diversity of the intestinal epithelium and are ideally suited for in situ visualization and continuous monitoring of epithelial development and differentiation 4–8. However, the presence of an enclosed lumen is non-physiological, as secreted material from goblet,
Gut-on-a-Chip microenvironment induces human intestinal cells to undergo villus differentiation
Integrative biology : quantitative biosciences from nano to macro, 2013
Existing in vitro models of human intestinal function commonly rely on use of established epithelial cell lines, such as Caco-2 cells, which form polarized epithelial monolayers but fail to mimic more complex intestinal functions that are required for drug development and disease research. We show here that a microfluidic 'Gut-on-a-Chip' technology that exposes cultured cells to physiological peristalsis-like motions and liquid flow can be used to induce human Caco-2 cells to spontaneously undergo robust morphogenesis of three-dimensional (3D) intestinal villi. The cells of that line these villus structures are linked by tight junctions, and covered by brush borders and mucus. They also reconstitute basal proliferative crypts that populate the villi along the crypt-villus axis, and form four different types of differentiated epithelial cells (absorptive, mucus-secretory, enteroendocrine, and Paneth) that take characteristic positions similar to those observed in living human...
Microfluidic Gut-on-a-Chip: Fundamentals and Challenges
Biosensors
The human gut is responsible for food digestion and absorption. Recently, growing evidence has shown its vital role in the proper functioning of other organs. Advances in microfluidic technologies have made a significant impact on the biomedical field. Specifically, organ-on-a-chip technology (OoC), which has become a popular substitute for animal models, is capable of imitating complex systems in vitro and has been used to study pathology and pharmacology. Over the past decade, reviews published focused more on the applications and prospects of gut-on-a-chip (GOC) technology, but the challenges and solutions to these limitations were often overlooked. In this review, we cover the physiology of the human gut and review the engineering approaches of GOC. Fundamentals of GOC models including materials and fabrication, cell types, stimuli and gut microbiota are thoroughly reviewed. We discuss the present GOC model applications, challenges, possible solutions and prospects for the GOC m...
A Micro-engineered Human Colon Intestine-Chip Platform to Study Leaky Barrier
bioRxiv (Cold Spring Harbor Laboratory), 2020
The intestinal epithelial barrier supports the symbiotic relationship between the microbiota colonizing the intestinal epithelium and the host immune system to maintain homeostasis. Leaky barrier is increasingly recognized as part of the pathogenesis of a number of chronic conditions in addition to inflammatory and infectious diseases. As our understanding on the regulation of the barrier remains limited, effective therapeutic targeting for the compromised barrier is still an unmet need. Here we combined advancements on the organoids and Organ-on-Chip technologies to establish a microengineered Colon Intestine-Chip for studying development and regulation of the human intestinal barrier. Our data demonstrate the significance of the endothelium in co-culture with the epithelial cells within a tissue-relevant microenvironment for the establishment of a tight epithelial barrier of polarized cells. Pathway analysis of the RNA sequencing (RNA-Seq), revealed significant upregulation of mechanisms relevant to the maturation of the intestinal epithelium in organoid-derived epithelial cells in co-culture with endothelium as compared to organoids maintained in suspension. We provide evidence that the Colon Intestine-Chip platform responds to interferon gamma (IFNγ), a prototype cytokine utilized to model inflammation-induced barrier disruption, by induction of apoptosis and reorganization of the apical junctional complexes as shown with other systems. We also describe the mechanism of action of interleukin 22 (IL-22) on mature, organoid-derived intestinal epithelial cells that is consistent with barrier disruption. Overall we propose the Colon Intestine-Chip as a promising human organoid-derived platform to decipher mechanisms driving the development of leaky gut in patients and enable their translation for this unmet medical need. .
In vitro models and ex vivo systems used in inflammatory bowel disease
In vitro models
Inflammatory bowel disease (IBD) is a chronic, relapsing gastrointestinal condition. Ulcerative colitis and Crohn's disease are types of inflammatory bowel disease. Over many decades, the disease has been a topic of study, with experts still trying to figure out its cause and pathology. Researchers have established many in vivo animal models, in vitro cell lines, and ex vivo systems to understand its cause ultimately and adequately identify a therapy. However, in vivo animal models cannot be regarded as good models for studying IBD since they cannot completely simulate the disease. Furthermore, because species differences are a crucial subject of concern, in vitro cell lines and ex vivo systems can be employed to recreate the condition properly. In vitro models serve as the starting point for biological and medical research. Ex vivo and in vitro models for replicating gut physiology have been developed. This review aims to present a clear understanding of several in vitro and ex vivo models of IBD and provide insights into their benefits and limits and their value in understanding intestinal physiology. Keywords Inflammatory bowel disease • In vitro cell lines • Ex vivo systems • Organoids • Caco-2 cell lines • Gut-on-chip