Gut like structures and gut tissues from mouse ES cells and their usage for developmental biology

该项目的目的是建立小鼠胚胎干细胞培养系统，以形成肠道样结构，作为体外发育模型，并用于肠道器官发生的研究。我们建立了培养体系，通过滴滴形成胚状体，并将其镀在明胶包覆的培养皿上生长。对肠样结构进行形态学和分子生物学分析。我们发现肠样结构的形成过程模拟了肠道器官发生，并及时表达了对肠道发育至关重要的转录因子。因此，我们得出结论，我们的系统是肠道发育的体外模型。我们在适当的期刊上发表了数据。胃肠道肌肉组织的分化机制尚不清楚，有研究认为骨形态发生蛋白（Bone Morphogenetic Protein， BMP）信号参与了这一过程。我们使用肠样结构来分析其机理，并将琼脂糖珠浸泡在形成肠样结构的附近。在肠样结构的平滑肌分化过程中，bmp尤其是BMP2促进了平滑肌的分化，形成了大量具有自发节律性收缩的肌肉组织。Real - time PCR结果显示，植入BMP微球后，一些与平滑肌分化相关的转录因子增加。我们正在处理数据准备发表。

The aims of the project were establishment of the culture system to develop gut-like structures from mouse ES cells as an in vitro developmental model and usage for investigation of gut organogensis. We established the culture system by hanging drops to form embryoid bodies and plating them on gelatin coated dishes to outgrowth. The gut-like structures were analyzed morphologically and molecular biologically. We found that the formation process of gut-like structures mimicked gut organogenesis and transcription factors crucial for the gut development were timely expressed. Therefore, we concluded our system was an in vitro model of gut development. We published data in appropriate journals.The differentiation mechanism of musculature of the gastrointestinal (GI) tract has been remained to clarify and it has been suggested Bone Morphogenetic Protein (BMP) signaling is involved in the process. We used gut-like structures for analysis of the mechanism and put agarose beads soaked with BMPs near developing gut-like structures. During the smooth muscle differentiation in the gut-like structures, BMPs especially BMP2 advanced the differentiation and formed substantial musculature showing spontaneous rhythmical contractions. Real time PCR revealed some transcription factors relating to smooth muscle differentiation increased after insertion of BMP beads. We are processing the data for publication now.