Long-term flow through human intestinal organoids with the gut organoid flow chip (GOFlowChip)

Sidar, Barkan; Jenkins, Brittany R.; Huang, Sha; Spence, Jason R.; Walk, Seth T.; Wilking, James N.

doi:10.1039/c9lc00653b

Cited by 61 publications

(45 citation statements)

References 88 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Intestinal organoids or enteroids have been proposed as an alternative model system to study the molecular aspects of host-pathogen interactions, especially as they apply to the gastrointestinal tract [44][45][46][51][52][53][54][55][56][57]. This approach is built on certain virtues that organotypic systems possess, including that they are untransformed and human, heterocellular [58][59][60], grow in three dimensions [46], ability to engineer "enhancements" such as co-culture with immune cells [61], flow in one direction [62], and vasculature [63] that undergo physiologic responses to stimuli such as fluid secretion [51, 64,65], mucus production [45], and innate immune responses [56,61]. In addition, researches could (i) assess all four intestinal segments from…”

Section: Discussionmentioning

confidence: 99%

Enteroaggregative E. coli Adherence to Human Heparan Sulfate Proteoglycans Drives Segment and Host Specific Responses to Infection

et al. 2020

View full text Add to dashboard Cite

Enteroaggregative Escherichia coli (EAEC) is a significant cause of acute and chronic diarrhea, foodborne outbreaks, infections of the immunocompromised, and growth stunting in children in developing nations. There is no vaccine and resistance to antibiotics is rising. Unlike related E. coli pathotypes that are often associated with acute bouts of infection, EAEC is associated with persistent diarrhea and subclinical long-term colonization. Several secreted virulence factors have been associated with EAEC pathogenesis and linked to disease in humans, less certain are the molecular drivers of adherence to the intestinal mucosa. We previously established human intestinal enteroids (HIEs) as a model system to study host-EAEC interactions and aggregative adherence fimbriae A (AafA) as a major driver of EAEC adherence to HIEs. Here, we report a large-scale assessment of the host response to EAEC adherence from all four segments of the intestine across at least three donor lines for five E. coli pathotypes. The data demonstrate that the host response in the duodenum is driven largely by the infecting pathotype, whereas the response in the colon diverges in a patient-specific manner. Major pathways altered in gene expression in each of the four enteroid segments differed dramatically, with responses observed for inflammation, apoptosis and an overwhelming response to different mucin genes. In particular, EAEC both associated with large mucus droplets and specific mucins at the epithelial surface, binding

show abstract

Section: Discussionmentioning

confidence: 99%

Enteroaggregative E. coli Adherence to Human Heparan Sulfate Proteoglycans Drives Segment and Host Specific Responses to Infection

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Alterations to the HIO culture system that enable co-differentiation or co-culture of endothelium [14], neuron [17], immune or other lineages are interesting approaches to achieve a more complete model of human intestine development, and may allow enhanced maturation of cell types. Furthermore, maturation may also require perfusable vasculature [74,75], extracellular matrix alterations [10], or other mechanical inductive cues [76]. Achieving mature intestinal tissue from iPSCs in vitro remains a major challenge in the field, and continued benchmarking against multi-organ reference atlases such as the one generated here, will be required to quantify the precision of cell state specification.…”

Section: Discussionmentioning

confidence: 99%

An organoid and multi-organ developmental cell atlas reveals multilineage fate specification in the human intestine

Kilik

Holloway

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Human intestinal organoids (HIOs) generated from pluripotent stem cells provide extraordinary opportunities to explore development and disease. Here, we generate a single-cell transcriptome reference atlas from HIOs and from multiple developing human organs to quantify the specificity of HIO cell fate acquisition, and to explore alternative fates. We identify epithelium-mesenchyme interactions, transcriptional regulators involved in cell fate specification, and stem cell maturation features in the primary tissue that are recapitulated in HIOs. We use an HIO time course to reconstruct the molecular dynamics of intestinal stem cell emergence, as well as the specification of multiple mesenchyme subtypes. We find that the intestinal master regulator CDX2 correlates with distinct phases of epithelial and mesenchymal development, and CDX2 deletion perturbs the differentiation of both intestinal epithelium and mesenchyme. Collectively our data provides a comprehensive and quantitative assessment of HIO development, and illuminates the molecular machinery underlying endodermal and mesodermal cell fate specification.

show abstract

“…We have shown that the HIO epithelium produces antimicrobial peptides, which could be particularly stressful to microbes within the small, static luminal space (14). To this point, since there is no peristalsis and luminal flow within HIOs, this leads to a buildup of epithelial and microbe-derived waste within the HIO lumen (28). In this regard, technological advancements that establish steady-state liquid flow through the HIO lumen (28) may lessen or eliminate microbial stressors within the HIO microenvironment.…”

Section: Discussionmentioning

confidence: 98%

The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: Escherichia coli Deficient in RpoS as a Colonization Probe

Barron

Cieza

Hill

et al. 2020

mSphere

Self Cite

View full text Add to dashboard Cite

Pluripotent stem-cell-derived human intestinal organoids (HIOs) are three-dimensional, multicellular structures that model a naive intestinal epithelium in an in vitro system. Several published reports have investigated the use of HIOs to study host-microbe interactions. We recently demonstrated that microinjection of the nonpathogenic Escherichia coli strain ECOR2 into HIOs induced morphological and functional maturation of the HIO epithelium, including increased secretion of mucins and cationic antimicrobial peptides. In the current work, we use ECOR2 as a biological probe to further characterize the environment present in the HIO lumen. We generated an isogenic mutant in the general stress response sigma factor RpoS and employed this mutant to compare challenges faced by a bacterium during colonization of the HIO lumen relative to the germ-free mouse intestine. We demonstrate that the loss of RpoS significantly decreases the ability of ECOR2 to colonize HIOs, although it does not prevent colonization of germ-free mice. These results indicate that the HIO lumen is a more restrictive environment to E. coli than the germ-free mouse intestine, thus increasing our understanding of the HIO model system as it pertains to studying the establishment of intestinal host-microbe symbioses. IMPORTANCE Technological advancements have driven and will continue to drive the adoption of organotypic systems for investigating host-microbe interactions within the human intestinal ecosystem. Using E. coli deficient in the RpoS-mediated general stress response, we demonstrate that the type or severity of microbial stressors within the HIO lumen is more restrictive than those of the in vivo environment of the germ-free mouse gut. This study provides important insight into the nature of the HIO microenvironment from a microbiological standpoint.

show abstract

Long-term flow through human intestinal organoids with the gut organoid flow chip (GOFlowChip)

Abstract: Human intestinal organoids (HIOs) have many research applications but are structurally limited with closed epithelial surfaces that limit material transport. Here, we present a fluidic device designed to establish long-term flow through HIOs.

Cited by 61 publications

References 88 publications

Enteroaggregative E. coli Adherence to Human Heparan Sulfate Proteoglycans Drives Segment and Host Specific Responses to Infection

Enteroaggregative E. coli Adherence to Human Heparan Sulfate Proteoglycans Drives Segment and Host Specific Responses to Infection

An organoid and multi-organ developmental cell atlas reveals multilineage fate specification in the human intestine

The Lumen of Human Intestinal Organoids Poses Greater Stress to Bacteria Compared to the Germ-Free Mouse Intestine: Escherichia coli Deficient in RpoS as a Colonization Probe

Contact Info

Product

Resources

About