Gut-like structures from mouse embryonic stem cells as an in vitro model for gut organogenesis preserving developmental potential after transplantation

¹ Department of Anatomy & Molecular Cell Biology, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya, Japan

^* To whom correspondence should be addressed. E-mail: storiha{at}met.nagoya-u.ac.jp.

	Abstract

Recently, we reported the formation of gut-like structures from mouse ES cells in vitro. To determine whether ES cells provide an in vitro model of gastrointestinal (GI) tracts and their organogenesis, we investigated the morphological features, formation process, cellular development and regional location within the GI tract by immunohistochemistry, electron microscopy and RT-PCR. We also examined the developmental potential by transplantation into kidney capsules. The results demonstrated that Id2 expressing epithelium developed first, -smooth muscle actin appeared around the periphery, and finally the gut-like structures were formed into three layer organ with well-differentiated epithelium. A connective tissue layer and musculature with interstitial cells of Cajal developed similar to organogenesis of the embryonic gut. Enteric neurons appeared underdeveloped and blood vessels were absent. Many structures expressed intestinal markers, Cdx2 and 5-HT, but not the stomach marker, H⁺/K⁺ ATPase. Transplantation obtained blood vessels and extrinsic nerve growth from the host to prolong life, and even grafts of premature structures did not form teratoma. In conclusion, gut-like structures were provided with prototypical tissue components of the GI tract and are inherent in the intestine rather than the stomach. The formation process was basically same as in gut organogenesis. They maintain their developmental potential after transplantation. Therefore gut-like structures provide a unique and useful in vitro system for development and stem cell studies of the GI tract including transplantation experiments.