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A Novel Method for the Culture and Polarized Stimulation of Human Intestinal Mucosa Explants
Open accessAbstract
Few models currently exist to realistically simulate the complex human intestine's micro-environment, where a variety of interactions take place. Proper homeostasis directly depends on these interactions, as they shape an entire immunological response inducing tolerance against food antigens while at the same time mounting effective immune responses against pathogenic microbes accidentally ingested with food. Intestinal homeostasis is preserved also through various complex interactions between the microbiota (including food-associated beneficial bacterial strains) and the host, that regulate the attachment/degradation of mucus, the production of antimicrobial peptides by the epithelial barrier, and the "education" of epithelial cells' that controls the tolerogenic or immunogenic phenotype of unique, gut-resident lymphoid cells' populations. These interactions have been so far very difficult to reproduce with in vitro assays using either cultured cell lines or peripheral blood mononuclear cells. In addition, mouse models differ substantially in components of the intestinal mucosa (mucus layer organization, commensal bacteria community) with respect to the human gut. Thus, studies of a variety of treatments to be brought in the clinics for important stress-related or pathological conditions such as irritable bowel syndrome, inflammatory bowel disease or colorectal cancer have been difficult to carry out. To address these issues, we developed a novel system that enables us to stimulate explants of human intestinal mucosa that retain their in situ conditioning by the host microbiota and immune response, in a polarized fashion. Polarized apical stimulation is of great importance for the outcome of the elicited immune response. It has been repeatedly shown that the same stimuli can produce completely different responses when they bypass the apical face of the intestinal epithelium, stimulating epithelial cells basolaterally or coming into direct contact with lamina propria components, switching the phenotype from tolerogenic to immunogenic and causing unnecessary and excessive inflammation in the area. We achieved polarized stimulation by gluing a cave cylinder which delimited the area of stimulation on the apical face of the mucosa as will be described in the protocol. We used this model to examine, among others, differential effects of three different Lactobacilli strains. We show that this model system is very powerful to assess the immunomodulatory properties of probiotics in healthy and disease conditions.
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In vitro intestine models created using human fetal intestinal tissue explants deteriorate within 24 hours of culture.
“In vitro intestine models have been created using human fetal intestinal tissue explants, but they deteriorate within 24 hours of culture.45, 46”
From Microfluidic Organ-on-a-Chip Models of Human Intestine (2018) · cites this paper as [45]
The Discussion describes a human intestinal mucosa explant model, but the tissue is obtained during surgery; it does not identify the explants as human fetal tissue. The Representative Results report that the explants did not survive 24 hours in a conventional incubator, but remained viable for at least 24 hours when cultured in oxygen. Thus, deterioration within 24 hours is supported only under a condition the claim omits. (LLM, conf=0.87)
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Development of a primary human Small Intestine-on-a-Chip using biopsy-derived organoids
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Metadata
- Authors
- Tsilingiri, K., Sonzogni, A., Caprioli, F., & Rescigno, M.
- Affiliations
- European Institute of Oncology (IT)
- Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico (IT)
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2013-05-01
- Issue
- 75
- Pages
- e4368-e4368
- ISSN
- 1940-087X (ISSN-L)
- DOI
- 10.3791/4368
- PMC
- PMC3667689
- OpenAlex
- W1967051954
- Cited by
- 17
- References
- 12
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 1 · 2021: 3 · 2020: 4 · 2019: 1 · 2018: 3 · 2017: 1 · 2016: 2
- FWCI
- 0.30
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Visualized Experiments (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Fondazione Cariplo
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
- Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
- Health Sciences › Medicine › Gastroenterology › Inflammatory Bowel Disease
- Keywords
- intestinal homeostasis, probiotics, immunomodulation, gut microbiota, intestinal epithelium, irritable bowel syndrome, inflammatory bowel disease, colorectal cancer, mucus layer, antimicrobial peptides
- MeSH terms
- Humans, Intestinal Mucosa, Lactobacillus, Salmonella, Cell Culture Techniques, Probiotics
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