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An organotypic slice model for ex vivo study of neural, immune, and microbial interactions of mouse intestine.
- Source :
-
American journal of physiology. Gastrointestinal and liver physiology [Am J Physiol Gastrointest Liver Physiol] 2016 Feb 15; Vol. 310 (4), pp. G240-8. Date of Electronic Publication: 2015 Dec 17. - Publication Year :
- 2016
-
Abstract
- Organotypic tissue slices provide seminatural, three-dimensional microenvironments for use in ex vivo study of specific organs and have advanced investigative capabilities compared with isolated cell cultures. Several characteristics of the gastrointestinal tract have made in vitro models for studying the intestine challenging, such as maintaining the intricate structure of microvilli, the intrinsic enteric nervous system, Peyer's patches, the microbiome, and the active contraction of gut muscles. In the present study, an organotypic intestinal slice model was developed that allows for functional investigation across regions of the intestine. Intestinal tissue slices were maintained ex vivo for several days in a physiologically relevant environment that preserved normal enterocyte structure, intact and proliferating crypt cells, submucosal organization, and muscle wall composure. Cell death was measured by a membrane-impermeable DNA binding indicator, ethidium homodimer, and less than 5% of cells were labeled in all regions of the villi and crypt epithelia at 24 h ex vivo. This tissue slice model demonstrated intact myenteric and submucosal neuronal plexuses and functional interstitial cells of Cajal to the extent that nonstimulated, segmental contractions occurred for up to 48 h ex vivo. To detect changes in physiological responses, slices were also assessed for segmental contractions in the presence and absence of antibiotic treatment, which resulted in slices with lesser or greater amounts of commensal bacteria, respectively. Segmental contractions were significantly greater in slices without antibiotics and increased native microbiota. This model renders mechanisms of neuroimmune-microbiome interactions in a complex gut environment available to direct observation and controlled perturbation.<br /> (Copyright © 2016 the American Physiological Society.)
- Subjects :
- Animals
Anti-Bacterial Agents pharmacology
Calcium Channel Blockers pharmacology
Cell Proliferation drug effects
Enterocytes drug effects
Enterocytes physiology
Enterocytes ultrastructure
Female
Intestinal Mucosa immunology
Intestinal Mucosa innervation
Intestinal Mucosa microbiology
Intestinal Mucosa physiology
Intestines microbiology
Male
Mice
Mice, Inbred C57BL
Microbiota
Models, Biological
Muscle Contraction drug effects
Muscle Contraction physiology
Muscle, Smooth physiology
Nicardipine pharmacology
Organ Culture Techniques
Peyer's Patches immunology
Peyer's Patches innervation
Peyer's Patches microbiology
Intestines immunology
Intestines innervation
Subjects
Details
- Language :
- English
- ISSN :
- 1522-1547
- Volume :
- 310
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- American journal of physiology. Gastrointestinal and liver physiology
- Publication Type :
- Academic Journal
- Accession number :
- 26680736
- Full Text :
- https://doi.org/10.1152/ajpgi.00299.2015