Crucial transcription factors in endoderm and embryonic gut development are expressed in gut-like structures from mouse ES cells

¹ Dept. of Anatomy & Molecular Cell Biology, Nagoya University Graduate School of Medicine, Nagoya, Japan
² Dept. of Anatomy, Graduate School of Agriculture & Life Science, University of Tokyo, Tokyo, Japan
³ Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Japan

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

	Abstract

Mouse embryonic stem cells (ES cells) are pluripotent and retain the potential to form an organ similar to gut showing spontaneous contractions in vitro. The morphological features and their formation process using the hanging drop method to compose embryoid bodies (EBs) seem to be similar to those in vivo. To determine whether the same molecular mechanisms are involved in the formation process, the expressions of transcription factors regulating endoderm and gut development in the mouse embryo were examined by in situ hybridization and compared to those in vivo. The expressions of gene products were also examined by immunohistochemistry and their co-localization was analyzed with double staining. The results showed that all factors examined, i.e., Sox17, Id2, HNF3/Foxa2, and GATA4, were expressed in both EBs and the gut-like structures. Moreover, their expression patterns were similar to those in the mouse embryo. EBs, after the hanging drop and before outgrowth, already expressed all factors co-localized with each other at the EB epithelial structures. These findings suggest that the origin of gut-like structure is determined and formed as the epithelial structure in EB during the hanging drop, and they also indicate that the in vitro system using mouse ES cells mimics the development in vivo and should prove useful in the study of molecular mechanisms for endoderm and gut development.

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