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

本项目的目的是建立小鼠胚胎干细胞体外发育模型的培养体系，并将其用于肠道器官发生的研究。我们建立了培养体系，通过悬挂液滴形成胚状体，并将它们涂在涂有明胶的培养皿上，使其长出。对肠样结构进行了形态和分子生物学分析。我们发现，模拟肠道器官发生的肠样结构的形成过程，对肠道发育至关重要的转录因子得到了及时的表达。因此，我们得出结论，我们的系统是一个肠道发育的体外模型。我们在相关杂志上发表了相关数据。胃肠道肌肉系统的分化机制仍未阐明，有研究表明骨形态发生蛋白(BMP)信号转导参与了这一过程。我们使用肠样结构来分析这种机制，并将浸泡了骨形态发生蛋白的琼脂糖珠放在发育中的肠样结构附近。在肠样结构的平滑肌分化过程中，BMPs尤其是BMP2促进了分化，并形成了大量的肌肉组织，表现出自发的节律性收缩。实时荧光定量聚合酶链式反应显示，植入BMP微珠后，一些与平滑肌分化相关的转录因子表达增加。我们现在正在处理数据以供发布。

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.