Coculture of primary human colon monolayer with human gut bacteria

Zhang, Jianbo; Hernandez-Gordillo, Victor; Trapečar, Martin; Wright, C. Wayne; Taketani, Mao; Schneider, Kirsten; Chen, Wen Li Kelly; Stas, Eric; Breault, David T.; Voigt, Christopher A.; Griffith, Linda G.

doi:10.1038/s41596-021-00562-w

Cited by 38 publications

(27 citation statements)

References 57 publications

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“…Altering the apical media formulation via the addition of 3 mM of the antioxidant N-acetyl cysteine (NAC), a common antioxidant used in primary intestinal cultures 43 – 46 , resulted in drastically reduced ROS concentration for all co-cultures over the course of the experiment. Maximum average ROS levels in the absence of NAC ranged from 4.4 to 3.4 mM across each of the four experimental groups.…”

Section: Resultsmentioning

confidence: 99%

Reactive oxygen species limit intestinal mucosa-bacteria homeostasis in vitro

Luchan

Choi

Carrier

2021

Sci Rep

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Interactions between epithelial and immune cells with the gut microbiota have wide-ranging effects on many aspects of human health. Therefore, there is value in developing in vitro models capable of performing highly controlled studies of such interactions. However, several critical factors that enable long term homeostasis between bacterial and mammalian cultures have yet to be established. In this study, we explored a model consisting of epithelial and immune cells, as well as four different bacterial species (Bacteroides fragilis KLE1958, Escherichia coli MG1655, Lactobacillus rhamnosus KLE2101, or Ruminococcus gnavus KLE1940), over a 50 hour culture period. Interestingly, both obligate and facultative anaerobes grew to similar extents in aerobic culture environments during the co-culture period, likely due to measured microaerobic oxygen levels near the apical surface of the epithelia. It was demonstrated that bacteria elicited reactive oxygen species (ROS) production, and that the resulting oxidative damage heavily contributed to observed epithelial barrier damage in these static cultures. Introduction of a ROS scavenger significantly mitigated oxidative damage, improving cell monolayer integrity and reducing lipid peroxidation, although not to control (bacteria-free culture) levels. These results indicate that monitoring and mitigating ROS accumulation and oxidative damage can enable longer term bacteria-intestinal epithelial cultures, while also highlighting the significance of additional factors that impact homeostasis in mammalian cell-bacteria systems.

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Section: Resultsmentioning

confidence: 99%

Reactive oxygen species limit intestinal mucosa-bacteria homeostasis in vitro

Luchan

Choi

Carrier

2021

Sci Rep

Self Cite

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“…They can be differentiated towards intestinal, 39 pancreatic ductlike, 227 liver 33 228 and gastric 223 Available techniques ⇒ Apical-out (polarity inversion, allows access to the apical but not basal site). 114 241 ⇒ Planar cultures: monolayer, cells adherent to plate (easy to perform, allows access to the apical but not basal site), [241][242][243] air-liquid Interface (epithelial injury responses, host-microbe interaction, allows access to both, the apical and basal site). 121 241 243 ⇒ Microinjection (technique equipment is needed, hostmicrobe interaction).…”

Section: Key Information and Available Techniquesmentioning

confidence: 99%

Organoids in gastrointestinal diseases: from experimental models to clinical translation

Günther

Winner

Stappenbeck

2022

Gut

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We are entering an era of medicine where increasingly sophisticated data will be obtained from patients to determine proper diagnosis, predict outcomes and direct therapies. We predict that the most valuable data will be produced by systems that are highly dynamic in both time and space. Three-dimensional (3D) organoids are poised to be such a highly valuable system for a variety of gastrointestinal (GI) diseases. In the lab, organoids have emerged as powerful systems to model molecular and cellular processes orchestrating natural and pathophysiological human tissue formation in remarkable detail. Preclinical studies have impressively demonstrated that these organs-in-a-dish can be used to model immunological, neoplastic, metabolic or infectious GI disorders by taking advantage of patient-derived material. Technological breakthroughs now allow to study cellular communication and molecular mechanisms of interorgan cross-talk in health and disease including communication along for example, the gut–brain axis or gut–liver axis. Despite considerable success in culturing classical 3D organoids from various parts of the GI tract, some challenges remain to develop these systems to best help patients. Novel platforms such as organ-on-a-chip, engineered biomimetic systems including engineered organoids, micromanufacturing, bioprinting and enhanced rigour and reproducibility will open improved avenues for tissue engineering, as well as regenerative and personalised medicine. This review will highlight some of the established methods and also some exciting novel perspectives on organoids in the fields of gastroenterology. At present, this field is poised to move forward and impact many currently intractable GI diseases in the form of novel diagnostics and therapeutics.

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“…Advances in coupling microfluidic and cell culture approaches with 3D organ-on-a-chip systems have allowed for novel experimental possibilities and have greatly increased in vitro model complexity ( 62 ). In recent work, leveraging a Caco-2 cell model, researchers investigated gut epithelial barrier integrity in the presence of different human commensal strains, highlighting the power of these more advanced in vitro systems ( 63 ).…”

Section: In Vitro Approachesmentioning

confidence: 99%

Perspective: Leveraging the Gut Microbiota to Predict Personalized Responses to Dietary, Prebiotic, and Probiotic Interventions

Gibbons

Gurry

Lampe

et al. 2022

Advances in Nutrition

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Humans often show variable responses to dietary, prebiotic, and probiotic interventions. Emerging evidence indicates that the gut microbiota is a key determinant for this population heterogeneity. Here, we provide an overview of some of the major computational and experimental tools being applied to critical questions of microbiota-mediated personalized nutrition and health. First, we discuss the latest advances in in silico modeling of the microbiota-nutrition-health axis, including the application of statistical, mechanistic, and hybrid artificial intelligence models. Second, we address high-throughput in vitro techniques for assessing inter-individual heterogeneity, from ex vivo batch culturing of stool and continuous culturing in anaerobic bioreactors, to more sophisticated organ-on-a-chip models that integrate both host and microbial compartments. Third, we explore in vivo approaches for better understanding personalized, microbiota-mediated responses to diet, prebiotics, and probiotics, from non-human animal models and human observational studies, to human feeding trials and crossover interventions. We highlight examples of existing, consumer-facing precision nutrition platforms that are currently leveraging the gut microbiota. Furthermore, we discuss how the integration of a broader set of the tools and techniques described in this piece can generate the data necessary to support a greater diversity of precision nutrition strategies. Finally, we present a vision of a precision nutrition and healthcare future, which leverages the gut microbiota to design effective, individual-specific interventions.

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Coculture of primary human colon monolayer with human gut bacteria

Cited by 38 publications

References 57 publications

Reactive oxygen species limit intestinal mucosa-bacteria homeostasis in vitro

Reactive oxygen species limit intestinal mucosa-bacteria homeostasis in vitro

Organoids in gastrointestinal diseases: from experimental models to clinical translation

Perspective: Leveraging the Gut Microbiota to Predict Personalized Responses to Dietary, Prebiotic, and Probiotic Interventions

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