In vitro development of gut-like tissue demonstrating rhythmic contractions from embryonic mouse intestinal cells.
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.
Ito-Dufros
Y
Y
Department of Pharmacology, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan. itoy@my-pharm.ac.jp
Funakoshi
Y
Y
Uehara
A
A
Oishi
K
K
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D007422:Intestines / Q000196:embryology / Q000502:physiology*; D051379:Mice; D008813:Mice, Inbred ICR; D009119:Muscle Contraction / Q000502:physiology*; D009130:Muscle, Smooth / Q000196:embryology / Q000502:physiology; D011247:Pregnancy; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043222:Ubiquitin Thiolesterase / Q000235:genetics
D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D019009:Proto-Oncogene Proteins c-kit; D043222:Ubiquitin Thiolesterase; C496976:Uchl1 protein, mouse; D002118:Calcium