In vitro development of gut-like tissue demonstrating rhythmic contractions from embryonic mouse intestinal cells
Y Ito-Dufros1, Y Funakoshi, A Uehara
1Department of Pharmacology, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan. itoy@my-pharm.ac.jp
Insights
Researchers developed a novel gut-like tissue model from embryonic mouse intestinal cells. This model exhibits rhythmic contractions, aiding the study of gastrointestinal motility development.
Area of Science:
- Developmental Biology
- Gastroenterology
- Tissue Engineering
Background:
- Intestinal rhythmic motility is crucial for digestion and is regulated by interstitial cells of Cajal (ICC) and the enteric nervous system.
- ICC are believed to differentiate from mesenchymal progenitor cells during late embryonic development, leading to rhythmic motility.
Purpose of the Study:
- To establish a culture system for reconstructing gut-like tissue from mid-embryonic mouse intestinal cells.
- To investigate the potential of this reconstructed tissue to exhibit rhythmic contractions and cellular network formation.
Main Methods:
- Dispersed cells from mid-embryonic mouse intestine were enzymatically isolated.
- Cells were cultured at high density in a collagen gel for one week to form gut-like tissue.
- Immunohistochemistry was used to detect neuronal and ICC marker proteins (PGP9.5, c-Kit).
- Spontaneous Ca(2+) oscillations in c-Kit-immunopositive cells were measured in the presence of nifedipine.
Main Results:
- The reconstituted gut-like tissue demonstrated rhythmic contractions.
- The tissue stained positive for PGP9.5 and c-Kit, indicating the presence of neurones and ICC.
- Network formation of nerve cells and ICC was observed within the cultured tissue.
- c-Kit-immunopositive cells exhibited spontaneous Ca(2+) oscillations, suggesting coupling to slow wave activity.
Conclusions:
- This culture system successfully reconstructs a functional gut-like tissue from embryonic intestinal cells.
- The model exhibits key cellular components and functional characteristics of the developing gastrointestinal system.
- This system provides a valuable tool for studying the developmental mechanisms underlying gastrointestinal motility.
Abstract:
The rhythmic motility of the intestine is regulated by the interstitial cells of Cajal (ICC) and the enteric nervous system. Rhythmic motility is considered to occur after the differentiation of mesenchymal progenitor cells into ICC during the late embryonic period. In this study, we successfully reconstructed a gut-like tissue demonstrating rhythmic contractions by culturing dispersed cells enzymatically isolated from the mouse intestine during the mid-embryonic period. These intestinal cells were reconstituted into a collagen gel at high density, made to proliferate considerably, and grew into a gut-like tissue after 1 week of culturing. The reconstituted tissue showed rhythmic contractions and stained positive for the specific marker proteins of neurones and ICC, PGP9.5 and c-Kit. The tissue also demonstrated network formation by developing nerve cells and ICC. Moreover, in the presence of nifedipine, c-Kit-immunopositive cells showed spontaneous Ca(2+) oscillation, which is considered to be coupled to the electrical activity that corresponds to slow waves. Therefore, this culture system may be of use in elucidating the developmental mechanisms of gastrointestinal motility.
